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Liang W, Liu Y, Guan H, Sakulsaengprapha V, Luby-Phelps K, Mahendroo M, Li X. Cervical Collagen Network Porosity Assessed by SHG Endomicroscopy Distinguishes Preterm and Normal Pregnancy-A Pilot Study. IEEE Trans Biomed Eng 2025; 72:777-785. [PMID: 39352817 PMCID: PMC11875912 DOI: 10.1109/tbme.2024.3472015] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/04/2024]
Abstract
OBJECTIVE Preterm birth (PTB) remains a pressing global health concern associated with premature cervical ripening and weakened cervical mechanical strength. Second harmonic generation (SHG) microscopy has proved instrumental in tracking progressive changes in cervical collagen morphology during pregnancy. To translate this imaging modality into clinical practice, we have developed a flexible SHG endomicroscope for label-free visualization of cervical collagen architecture. This study aims to assess the feasibility of our SHG endomicroscope for non-invasive differentiation of normal and PTB mouse models, with the ultimate goal of enabling early diagnosis and risk assessment of PTB in vivo. METHODS in this pilot investigation, we conducted endomicroscopic SHG imaging on frozen cervical tissue sections and intact cervices resected from both normal pregnant mice and mifepristone-induced PTB mouse models, and then analyzed the acquired images to identify collagen morphology characteristics associated with abnormal cervical collagen remodeling. RESULTS quantitative image analysis revealed significantly altered collage spatial distribution, larger collagen fiber diameter and pore size, along with reduced pore numbers in SHG endomicroscopy images from PTB mouse models compared to normal pregnant mice. Similar trends were consistent across SHG endomicroscopy images of subepithelial collagen fibers acquired directly from intact cervices. CONCLUSION/SIGNIFICANCE overall, the experiment results underscore the potential of SHG endomicroscopy, coupled with quantitative image analysis, for clinically evaluating cervical collagen remodeling and PTB risk.
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2
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Morton RA, Kim TN. Viscocohesive hyaluronan gel enhances stability of intravital multiphoton imaging with subcellular resolution. NEUROPHOTONICS 2025; 12:S14602. [PMID: 39583344 PMCID: PMC11582905 DOI: 10.1117/1.nph.12.s1.s14602] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Subscribe] [Scholar Register] [Received: 09/03/2024] [Revised: 10/29/2024] [Accepted: 11/06/2024] [Indexed: 11/26/2024]
Abstract
Multiphoton microscopy (MPM) has become a preferred technique for intravital imaging deep in living tissues with subcellular detail, where resolution and working depths are typically optimized utilizing high numerical aperture, water-immersion objectives with long focusing distances. However, this approach requires the maintenance of water between the specimen and the objective lens, which can be challenging or impossible for many intravital preparations with complex tissues and spatial arrangements. We introduce the novel use of cohesive hyaluronan gel (HG) as an immersion medium that can be used in place of water within existing optical setups to enable multiphoton imaging with equivalent quality and far superior stability. We characterize and compare imaging performance, longevity, and feasibility of preparations in various configurations. This combination of HG with MPM is highly accessible and opens the doors to new intravital imaging applications.
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Affiliation(s)
- Ryan A. Morton
- University of California San Francisco, Department of Ophthalmology, San Francisco, California, United States
| | - Tyson N. Kim
- University of California San Francisco, Department of Ophthalmology, San Francisco, California, United States
- UCSF-UC Berkeley Graduate Group in Bioengineering, San Francisco/Berkeley, California, United States
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3
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Zhang K, Ducourthial G, Sharan T, Mohan A, Hou J, Timilsina S, Liang R, Louradour F, Spring BQ. Video-rate two-photon microendoscopy using second harmonic resonance fiber scanning. BIOMEDICAL OPTICS EXPRESS 2024; 15:6324-6339. [PMID: 39553867 PMCID: PMC11563328 DOI: 10.1364/boe.534399] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Figures] [Subscribe] [Scholar Register] [Received: 07/08/2024] [Revised: 09/12/2024] [Accepted: 09/17/2024] [Indexed: 11/19/2024]
Abstract
We present a 2.5-mm-diameter resonant fiber scanning two-photon microendoscope with a 30-mm long forward-viewing rigid probe tip that enables video-rate imaging (20 Hz frame rate) suitable for hand-held imaging of tissues without motion artifacts. Higher-order harmonic oscillation scanning techniques are developed to significantly increase the frame rate compared to prior published fiber scanning microendoscopy designs while maintaining the field-of-view (∼125 µm), the optical resolution (1.2 µm lateral and 10.9 µm axial resolution, full width at half maximum), and the spatial sampling (1250 circumferential pixels per spiral × 20 radial pixels over the diameter; 210 spirals per frame, ∼4 spiral samples per resolvable pixel) compared to a traditional scan using the fundamental resonance. 3D printed mounts were created to reduce the cost and simplify the fabrication for the fiber scanner without compromising performance or stability (<0.3 µm drift over 84 hours). A custom long-wavelength (∼1.08 µm) femtosecond fiber laser is coupled into several meters of fiber to realize a flexible, hand-held device for long-wavelength multiphoton microendoscopy.
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Affiliation(s)
- Kai Zhang
- Department of Physics, Northeastern University, Boston, MA 02115, USA
- Translational Biophotonics Cluster, Northeastern University, Boston, MA 02115, USA
| | | | - Taresh Sharan
- Department of Physics, Northeastern University, Boston, MA 02115, USA
- Translational Biophotonics Cluster, Northeastern University, Boston, MA 02115, USA
- School of Biomedical Engineering, Indian Institute of Technology (BHU), Varanasi, Uttar Pradesh 221005, India
| | - Arvind Mohan
- Translational Biophotonics Cluster, Northeastern University, Boston, MA 02115, USA
- Department of Bioengineering, Northeastern University, Boston, MA 02115, USA
| | - Jiasen Hou
- Department of Physics, Northeastern University, Boston, MA 02115, USA
- Translational Biophotonics Cluster, Northeastern University, Boston, MA 02115, USA
| | - Sudip Timilsina
- Department of Physics, Northeastern University, Boston, MA 02115, USA
- Translational Biophotonics Cluster, Northeastern University, Boston, MA 02115, USA
| | - Rongguang Liang
- Wyant College of Optical Sciences, The University of Arizona, Tucson, AZ 85721, USA
| | | | - Bryan Q. Spring
- Department of Physics, Northeastern University, Boston, MA 02115, USA
- Translational Biophotonics Cluster, Northeastern University, Boston, MA 02115, USA
- Department of Bioengineering, Northeastern University, Boston, MA 02115, USA
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Ayubi GA, Dubra A. Robust real-time estimation of non-uniform angular velocity and sub-pixel jitter in images captured with resonant scanners. OPTICS EXPRESS 2023; 31:44199-44211. [PMID: 38178497 DOI: 10.1364/oe.512233] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 11/16/2023] [Accepted: 12/05/2023] [Indexed: 01/06/2024]
Abstract
Images captured with resonant scanners are affected by angular velocity fluctuations that result in image distortion and by poor synchronization between scanning and light detection that creates jitter between image rows. We previously demonstrated that both problems can be mitigated in post-processing by recording the scanner orientation in synchrony with the image capture, followed by data resampling [Opt. Express30, 112 (2022)10.1364/OE.446162]. Here we introduce more robust algorithms for estimation of both angular velocity fluctuation and jitter in the presence of random and deterministic noise. We also show linearization of the scanner oscillation model to reduce calculation times by two orders of magnitude, reaching 65,000 jitter estimations per second when using 2,800 samples per image row, and 500,000 when using only 500 samples, easily supporting real-time generation of jitter-corrected images.
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5
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Bae H, Rodewald M, Meyer-Zedler T, Bocklitz TW, Matz G, Messerschmidt B, Press AT, Bauer M, Guntinas-Lichius O, Stallmach A, Schmitt M, Popp J. Feasibility studies of multimodal nonlinear endoscopy using multicore fiber bundles for remote scanning from tissue sections to bulk organs. Sci Rep 2023; 13:13779. [PMID: 37612362 PMCID: PMC10447453 DOI: 10.1038/s41598-023-40944-6] [Citation(s) in RCA: 2] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/08/2023] [Accepted: 08/18/2023] [Indexed: 08/25/2023] Open
Abstract
Here, we report on the development and application of a compact multi-core fiber optical probe for multimodal non-linear imaging, combining the label-free modalities of Coherent Anti-Stokes Raman Scattering, Second Harmonic Generation, and Two-Photon Excited Fluorescence. Probes of this multi-core fiber design avoid moving and voltage-carrying parts at the distal end, thus providing promising improved compatibility with clinical requirements over competing implementations. The performance characteristics of the probe are established using thin cryo-sections and artificial targets before the applicability to clinically relevant samples is evaluated using ex vivo bulk human and porcine intestine tissues. After image reconstruction to counteract the data's inherently pixelated nature, the recorded images show high image quality and morpho-chemical conformity on the tissue level compared to multimodal non-linear images obtained with a laser-scanning microscope using a standard microscope objective. Furthermore, a simple yet effective reconstruction procedure is presented and demonstrated to yield satisfactory results. Finally, a clear pathway for further developments to facilitate a translation of the multimodal fiber probe into real-world clinical evaluation and application is outlined.
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Affiliation(s)
- Hyeonsoo Bae
- Leibniz Institute of Photonic Technology (Leibniz IPHT), Member of Leibniz Health Technologies, Member of the Leibniz Centre for Photonics in Infection Research (LPI), PO Box 100239, 07702, Jena, Germany
- Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich-Schiller University Jena, Helmholtzweg 4, 07743, Jena, Germany
- Center for Sepsis Control and Care (CSCC), Jena University Hospital, Erlanger Allee 101, 07747, Jena, Germany
| | - Marko Rodewald
- Leibniz Institute of Photonic Technology (Leibniz IPHT), Member of Leibniz Health Technologies, Member of the Leibniz Centre for Photonics in Infection Research (LPI), PO Box 100239, 07702, Jena, Germany
- Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich-Schiller University Jena, Helmholtzweg 4, 07743, Jena, Germany
| | - Tobias Meyer-Zedler
- Leibniz Institute of Photonic Technology (Leibniz IPHT), Member of Leibniz Health Technologies, Member of the Leibniz Centre for Photonics in Infection Research (LPI), PO Box 100239, 07702, Jena, Germany
- Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich-Schiller University Jena, Helmholtzweg 4, 07743, Jena, Germany
| | - Thomas W Bocklitz
- Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich-Schiller University Jena, Helmholtzweg 4, 07743, Jena, Germany
| | - Gregor Matz
- GRINTECH GmbH, Schillerstraße 1, 07745, Jena, Germany
| | | | - Adrian T Press
- Center for Sepsis Control and Care (CSCC), Jena University Hospital, Erlanger Allee 101, 07747, Jena, Germany
- Department of Anesthesiology and Intensive Care Medicine, Jena University Hospital, Am Klinikum 1, 07747, Jena, Germany
- Medical Faculty, Friedrich-Schiller University Jena, Kastanienstr. 1, 07747, Jena, Germany
| | - Michael Bauer
- Center for Sepsis Control and Care (CSCC), Jena University Hospital, Erlanger Allee 101, 07747, Jena, Germany
- Department of Anesthesiology and Intensive Care Medicine, Jena University Hospital, Am Klinikum 1, 07747, Jena, Germany
| | - Orlando Guntinas-Lichius
- Department of Otorhinolaryngology, Jena University Hospital, Am Klinikum 1, 07747, Jena, Germany
| | - Andreas Stallmach
- Department of Internal Medicine IV, Jena University Hospital, Am Klinikum 1, 07747, Jena, Germany
| | - Michael Schmitt
- Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich-Schiller University Jena, Helmholtzweg 4, 07743, Jena, Germany
| | - Juergen Popp
- Leibniz Institute of Photonic Technology (Leibniz IPHT), Member of Leibniz Health Technologies, Member of the Leibniz Centre for Photonics in Infection Research (LPI), PO Box 100239, 07702, Jena, Germany.
- Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich-Schiller University Jena, Helmholtzweg 4, 07743, Jena, Germany.
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Stanciu SG, König K, Song YM, Wolf L, Charitidis CA, Bianchini P, Goetz M. Toward next-generation endoscopes integrating biomimetic video systems, nonlinear optical microscopy, and deep learning. BIOPHYSICS REVIEWS 2023; 4:021307. [PMID: 38510341 PMCID: PMC10903409 DOI: 10.1063/5.0133027] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Received: 11/15/2022] [Accepted: 05/26/2023] [Indexed: 03/22/2024]
Abstract
According to the World Health Organization, the proportion of the world's population over 60 years will approximately double by 2050. This progressive increase in the elderly population will lead to a dramatic growth of age-related diseases, resulting in tremendous pressure on the sustainability of healthcare systems globally. In this context, finding more efficient ways to address cancers, a set of diseases whose incidence is correlated with age, is of utmost importance. Prevention of cancers to decrease morbidity relies on the identification of precursor lesions before the onset of the disease, or at least diagnosis at an early stage. In this article, after briefly discussing some of the most prominent endoscopic approaches for gastric cancer diagnostics, we review relevant progress in three emerging technologies that have significant potential to play pivotal roles in next-generation endoscopy systems: biomimetic vision (with special focus on compound eye cameras), non-linear optical microscopies, and Deep Learning. Such systems are urgently needed to enhance the three major steps required for the successful diagnostics of gastrointestinal cancers: detection, characterization, and confirmation of suspicious lesions. In the final part, we discuss challenges that lie en route to translating these technologies to next-generation endoscopes that could enhance gastrointestinal imaging, and depict a possible configuration of a system capable of (i) biomimetic endoscopic vision enabling easier detection of lesions, (ii) label-free in vivo tissue characterization, and (iii) intelligently automated gastrointestinal cancer diagnostic.
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Affiliation(s)
- Stefan G. Stanciu
- Center for Microscopy-Microanalysis and Information Processing, University Politehnica of Bucharest, Bucharest, Romania
| | | | | | - Lior Wolf
- School of Computer Science, Tel Aviv University, Tel-Aviv, Israel
| | - Costas A. Charitidis
- Research Lab of Advanced, Composite, Nano-Materials and Nanotechnology, School of Chemical Engineering, National Technical University of Athens, Athens, Greece
| | - Paolo Bianchini
- Nanoscopy and NIC@IIT, Italian Institute of Technology, Genoa, Italy
| | - Martin Goetz
- Medizinische Klinik IV-Gastroenterologie/Onkologie, Kliniken Böblingen, Klinikverbund Südwest, Böblingen, Germany
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7
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Liverman S, Trotter CW, Bouchard L, Bialek H, Nguyen T, Natarajan A, Wang AX. Dynamic indoor free-space optical communication enabled by beam steering and beam shaping. APPLIED OPTICS 2023; 62:2367-2375. [PMID: 37132876 DOI: 10.1364/ao.451422] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/04/2023]
Abstract
Indoor free-space optical communication (FSO) provides orders of magnitude larger usable bandwidth compared to radio-frequency links but suffers from an intrinsic trade-off between areal coverage and received power. In this paper, we report a dynamic indoor FSO system enabled by a line-of-sight optical link featuring advanced beam control capabilities. The optical link herein utilizes a passive target acquisition scheme by combining a beam steering and beam shaping transmitter with a receiver adorned with a ring-shaped retroreflector. When controlled by an efficient beam scanning algorithm, the transmitter is capable of locating the receiver with millimeter-scale accuracy over a distance of 3 m with a full viewing angle of ±11.25∘ in the vertical direction and ±18.75∘ in the horizontal direction within 1.162±0.005s, regardless of the receiver's positions. We also demonstrate 1 Gbit/s data rate with bit error rates below 4×10-7 using an 850 nm laser diode with only 2 mW of output power.
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8
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He L, Wen Z, Wang B, Li X, Wu D. Structural Design and Experimental Studies of Resonant Fiber Optic Scanner Driven by Co-Fired Multilayer Piezoelectric Ceramics. MICROMACHINES 2023; 14:517. [PMID: 36984924 PMCID: PMC10055889 DOI: 10.3390/mi14030517] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Subscribe] [Scholar Register] [Received: 01/24/2023] [Revised: 02/16/2023] [Accepted: 02/20/2023] [Indexed: 06/18/2023]
Abstract
Piezo-driven resonant fiber optic scanners are gaining more and more attention due to their simple structure, weak electromagnetic radiation, and non-friction loss. Conventional piezo-driven resonant fiber optic scanners typically use quadrature piezoelectric tubes (piezo tubes) operating in 31-mode with high drive voltage and low excitation efficiency. In order to solve the abovementioned problem, a resonant fiber scanner driven by co-fired multilayer piezoelectric ceramics (CMPCs) is proposed in which four CMPCs drive a cantilevered fiber optic in the first-order bending mode to achieve efficient and fast space-filling scanning. In this paper, the cantilever beam vibration model with base displacement excitation was derived to provide a theoretical basis for the design of the fiber optic scanner. The finite element method was used to guide the dynamic design of the scanner. Finally, the dynamics characteristics and scanning trajectory of the prepared scanner prototype were tested and compared with the theoretical and simulation calculation results. Experimental results showed that the scanner can achieve three types of space-filling scanning: spiral, Lissajous, and propeller. Compared with the structure using piezo tubes, the designed scanner achieved the same scanning range with smaller axial dimensions, lower drive voltage, and higher efficiency. The scanner can achieve a free end displacement of 10 mm in both horizontal and vertical directions under a sinusoidal excitation signal of 50 Vp-p and 200 Hz. The theoretical, simulation and experimental results validate the feasibility of the proposed scanner structure and provide new ideas for the design of resonant fiber optic scanners.
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9
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Villarreal PP, Pal R, Qiu S, Coblens O, Villasante-Tezanos A, Resto V, McCammon S, Vargas G. Label-Free Imaging and Histo-Optical Evaluation of Head and Neck Cancers with Multiphoton Autofluorescence Microscopy. Cancers (Basel) 2023; 15:1302. [PMID: 36831646 PMCID: PMC9953923 DOI: 10.3390/cancers15041302] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/07/2022] [Revised: 01/30/2023] [Accepted: 02/15/2023] [Indexed: 02/22/2023] Open
Abstract
Depth-resolved label-free optical imaging by the method of multiphoton autofluorescence microscopy (MPAM) may offer new ways to examine cellular and extracellular atypia associated with epithelial squamous cell carcinoma (SCC). MPAM was evaluated for its ability to identify cellular and microstructural atypia in head and neck tissues from resected discarded tumor tissue. Three-dimensional image volumes were obtained from tissues from the floor of the mouth, tongue, and larynx, and were then processed for histology. MPAM micrographs were evaluated for qualitative metrics of cell atypia and quantitative measures associated with nuclear pleomorphism. Statistical analyses correlated MPAM endpoints with histological grade from each imaged site. Cellular overcrowding, discohesion, anisonucleosis, and multinucleated cells, as observed through MPAM, were found to be statistically associated with dysplasia and SCC grading, but not in histologically benign regions. A quantitative measure of the coefficient of variance in nuclear size in SCC and dysplasia was statistically elevated above histologically benign regions. MPAM also allowed for the identification of cellular heterogeneity across transitional areas and other features, such as inflammatory infiltrates. In the future, MPAM could be evaluated for the non-invasive detection of neoplasia, possibly as an adjunct to traditional conventional examination and biopsy.
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Affiliation(s)
- Paula Patricia Villarreal
- The Institute for Translational Sciences, University of Texas Medical Branch, Galveston, TX 77555, USA
| | - Rahul Pal
- Athinoula A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital, Charlestown, MA 02129, USA
| | - Suimin Qiu
- Department of Pathology, Division of Surgical Pathology, University of Texas Medical Branch, Galveston, TX 77555, USA
| | - Orly Coblens
- Department of Otolaryngology, Head & Neck Surgery, University of Texas Medical Branch, Galveston, TX 77555, USA
| | - Alejandro Villasante-Tezanos
- Department of Biostatistics and Data Science, School for Public and Population Health, University of Texas Medical Branch, Galveston, TX 77555, USA
| | - Vicente Resto
- Department of Otolaryngology, Head & Neck Surgery, University of Texas Medical Branch, Galveston, TX 77555, USA
| | - Susan McCammon
- Department of Otolaryngology, Head & Neck Surgery Oncology Division, The University of Alabama at Birmingham, Birmingham, AL 35294, USA
| | - Gracie Vargas
- Department of Neurobiology, University of Texas Medical Branch, Galveston, TX 77555, USA
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10
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Kučikas V, Werner MP, Schmitz-Rode T, Louradour F, van Zandvoort MAMJ. Two-Photon Endoscopy: State of the Art and Perspectives. Mol Imaging Biol 2023; 25:3-17. [PMID: 34779969 PMCID: PMC9971078 DOI: 10.1007/s11307-021-01665-2] [Citation(s) in RCA: 19] [Impact Index Per Article: 9.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/18/2021] [Revised: 09/15/2021] [Accepted: 10/05/2021] [Indexed: 10/19/2022]
Abstract
In recent years, the demand for non-destructive deep-tissue imaging modalities has led to interest in multiphoton endoscopy. In contrast to bench top systems, multiphoton endoscopy enables subcellular resolution imaging in areas not reachable before. Several groups have recently presented their development towards the goal of producing user friendly plug and play system, which could be used in biological research and, potentially, clinical applications. We first present the technological challenges, prerequisites, and solutions in two-photon endoscopic systems. Secondly, we focus on the applications already found in literature. These applications mostly serve as a quality check of the built system, but do not answer a specific biomedical research question. Therefore, in the last part, we will describe our vision on the enormous potential applicability of adult two-photon endoscopic systems in biological and clinical research. We will thus bring forward the concept that two-photon endoscopy is a sine qua non in bringing this technique to the forefront in clinical applications.
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Affiliation(s)
- Vytautas Kučikas
- Institute for Molecular Cardiovascular Research (IMCAR), RWTH Aachen University, Aachen, Germany. .,XLIM Research Institute, Limoges University, CNRS, Limoges, France.
| | - Maximilian P Werner
- Department of Biohybrid and Medical Textiles (BioTex), RWTH Aachen University, Aachen, Germany
| | - Thomas Schmitz-Rode
- Department of Biohybrid and Medical Textiles (BioTex), RWTH Aachen University, Aachen, Germany
| | | | - Marc A M J van Zandvoort
- Institute for Molecular Cardiovascular Research (IMCAR), RWTH Aachen University, Aachen, Germany.,Institute for Cardiovascular Diseases CARIM, Department of Molecular Cell Biology, Maastricht University, Maastricht, Netherlands
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11
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Kreiss L, Ganzleben I, Mühlberg A, Ritter P, Schneidereit D, Becker C, Neurath MF, Friedrich O, Schürmann S, Waldner M. Label-free analysis of inflammatory tissue remodeling in murine lung tissue based on multiphoton microscopy, Raman spectroscopy and machine learning. JOURNAL OF BIOPHOTONICS 2022; 15:e202200073. [PMID: 35611635 DOI: 10.1002/jbio.202200073] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 03/16/2022] [Revised: 05/19/2022] [Accepted: 05/20/2022] [Indexed: 06/15/2023]
Abstract
Inflammatory fibrotic tissue remodeling can lead to severe morbidity. Histopathology grading requires extraction of biopsies and elaborate tissue processing. Label-free optical technologies can provide diagnostic readout without preparation and artificial stainings and show potential for in vivo applications. Here, we present an integration of Raman spectroscopy (RS) and multiphoton microscopy for joint investigation of the bio-chemical composition and morphological features related to cellular components and connective tissue. Both modalities show that collagen signatures were significantly increased in a murine fibrosis model. Furthermore, autofluorescence signatures assigned to immune cells show high correlation with disease severity. RS indicates increased levels of elastin and lipids. Further, we investigated the effect of joint data sets on prediction performance in machine learning models. Although binary classification did not benefit from adding more features, multi-class classification was improved by integrated data sets.
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Affiliation(s)
- Lucas Kreiss
- Institute of Medical Biotechnology, Friedrich-Alexander University Erlangen-Nürnberg, Erlangen, Germany
- Department of Medicine 1, University Hospital, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany
- Erlangen Graduate School in Advanced Optical Technologies (SAOT), Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany
| | - Ingo Ganzleben
- Department of Medicine 1, University Hospital, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany
- Ludwig Demling Center for Molecular Imaging, Department of Medicine 1, University Hospital, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany
| | - Alexander Mühlberg
- Institute of Medical Biotechnology, Friedrich-Alexander University Erlangen-Nürnberg, Erlangen, Germany
| | - Paul Ritter
- Institute of Medical Biotechnology, Friedrich-Alexander University Erlangen-Nürnberg, Erlangen, Germany
- Erlangen Graduate School in Advanced Optical Technologies (SAOT), Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany
| | - Dominik Schneidereit
- Institute of Medical Biotechnology, Friedrich-Alexander University Erlangen-Nürnberg, Erlangen, Germany
- Erlangen Graduate School in Advanced Optical Technologies (SAOT), Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany
| | - Christoph Becker
- Department of Medicine 1, University Hospital, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany
| | - Markus F Neurath
- Department of Medicine 1, University Hospital, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany
- Erlangen Graduate School in Advanced Optical Technologies (SAOT), Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany
- Ludwig Demling Center for Molecular Imaging, Department of Medicine 1, University Hospital, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany
| | - Oliver Friedrich
- Institute of Medical Biotechnology, Friedrich-Alexander University Erlangen-Nürnberg, Erlangen, Germany
- Erlangen Graduate School in Advanced Optical Technologies (SAOT), Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany
| | - Sebastian Schürmann
- Institute of Medical Biotechnology, Friedrich-Alexander University Erlangen-Nürnberg, Erlangen, Germany
- Erlangen Graduate School in Advanced Optical Technologies (SAOT), Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany
| | - Maximilian Waldner
- Department of Medicine 1, University Hospital, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany
- Erlangen Graduate School in Advanced Optical Technologies (SAOT), Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany
- Ludwig Demling Center for Molecular Imaging, Department of Medicine 1, University Hospital, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany
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12
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Choi W, Kang M, Hong JH, Katz O, Lee B, Kim GH, Choi Y, Choi W. Flexible-type ultrathin holographic endoscope for microscopic imaging of unstained biological tissues. Nat Commun 2022; 13:4469. [PMID: 35918348 PMCID: PMC9345988 DOI: 10.1038/s41467-022-32114-5] [Citation(s) in RCA: 12] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/16/2022] [Accepted: 07/18/2022] [Indexed: 12/20/2022] Open
Abstract
Ultrathin lensless fibre endoscopes offer minimally invasive investigation, but they mostly operate as a rigid type due to the need for prior calibration of a fibre probe. Furthermore, most implementations work in fluorescence mode rather than label-free imaging mode, making them unsuitable for general medical diagnosis. Herein, we report a fully flexible ultrathin fibre endoscope taking 3D holographic images of unstained tissues with 0.85-μm spatial resolution. Using a bare fibre bundle as thin as 200-μm diameter, we design a lensless Fourier holographic imaging configuration to selectively detect weak reflections from biological tissues, a critical step for label-free endoscopic reflectance imaging. A unique algorithm is developed for calibration-free holographic image reconstruction, allowing us to image through a narrow and curved passage regardless of fibre bending. We demonstrate endoscopic reflectance imaging of unstained rat intestine tissues that are completely invisible to conventional endoscopes. The proposed endoscope will expedite a more accurate and earlier diagnosis than before with minimal complications.
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Affiliation(s)
- Wonjun Choi
- Center for Molecular Spectroscopy and Dynamics, Institute for Basic Science, Seoul, Republic of Korea.,Department of Physics, Korea University, Seoul, Republic of Korea
| | - Munkyu Kang
- Center for Molecular Spectroscopy and Dynamics, Institute for Basic Science, Seoul, Republic of Korea.,Department of Physics, Korea University, Seoul, Republic of Korea
| | - Jin Hee Hong
- Center for Molecular Spectroscopy and Dynamics, Institute for Basic Science, Seoul, Republic of Korea.,Department of Physics, Korea University, Seoul, Republic of Korea
| | - Ori Katz
- Department of Applied Physics, The Selim and Rachel Benin School of Computer Science & Engineering, The Hebrew University of Jerusalem, Jerusalem, Israel
| | - Byunghak Lee
- Korea Electrotechnology Research Institute, Ansan, Korea.,B2LAB co., ltd, Pohang-si, Gyeongsangbuk, Korea
| | - Guang Hoon Kim
- Korea Electrotechnology Research Institute, Ansan, Korea
| | - Youngwoon Choi
- Department of Bioengineering, Korea University, Seoul, Republic of Korea. .,Interdisciplinary Program in Precision Public Health, Korea University, Seoul, Republic of Korea.
| | - Wonshik Choi
- Center for Molecular Spectroscopy and Dynamics, Institute for Basic Science, Seoul, Republic of Korea. .,Department of Physics, Korea University, Seoul, Republic of Korea.
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13
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Wang C, Liu H, Cui H, Ma J, Li Y, Tian J, Jin C, Chen Y, Gao Y, Fu Q, Hu Y, Wu D, Yu F, Wu R, Wang A, Feng L. Two-photon endomicroscopy with microsphere-spliced double-cladding antiresonant fiber for resolution enhancement. OPTICS EXPRESS 2022; 30:26090-26101. [PMID: 36236806 DOI: 10.1364/oe.461325] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 04/17/2022] [Accepted: 06/16/2022] [Indexed: 06/16/2023]
Abstract
We demonstrate a miniature fiber-optic two two-photon endomicroscopy with microsphere-spliced double-cladding antiresonant fiber for resolution enhancement. An easy-to-operate process for fixing microsphere permanently in an antiresonant fiber core, by arc discharge, is proposed. The flexible fiber-optic probe is integrated with a parameter of 5.8 mm × 49.1 mm (outer diameter × rigid length); the field of view is 210 µm, the resolution is 1.3 µm, and the frame rate is 0.7 fps. The imaging ability is verified using ex-vivo mouse kidney, heart, stomach, tail tendon, and in-vivo brain neural imaging.
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14
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Kreiss L, Thoma OM, Lemire S, Lechner K, Carlé B, Dilipkumar A, Kunert T, Scheibe K, Heichler C, Merten AL, Weigmann B, Neufert C, Hildner K, Vieth M, Neurath MF, Friedrich O, Schürmann S, Waldner MJ. Label-Free Characterization and Quantification of Mucosal Inflammation in Common Murine Colitis Models With Multiphoton Imaging. Inflamm Bowel Dis 2022; 28:1637-1646. [PMID: 35699622 PMCID: PMC9629455 DOI: 10.1093/ibd/izac114] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 12/30/2021] [Indexed: 12/13/2022]
Abstract
BACKGROUND Clinical challenges in inflammatory bowel diseases require microscopic in vivo evaluation of inflammation. Here, label-free imaging holds great potential, and recently, our group demonstrated the advantage of using in vivo multiphoton endomicroscopy for longitudinal animal studies. This article extends our previous work by in-depth analysis of label-free tissue features in common colitis models quantified by the multiphoton colitis score (MCS). METHODS Fresh mucosal tissues were evaluated from acute and chronic dextran sulfate sodium (DSS), TNBS, oxazolone, and transfer colitis. Label-free imaging was performed by using second harmonic generation and natural autofluorescence. Morphological changes in mucosal crypts, collagen fibers, and cellularity in the stroma were analyzed and graded. RESULTS Our approach discriminated between healthy (mean MCS = 2.5) and inflamed tissue (mean MCS > 5) in all models, and the MCS was validated by hematoxylin and eosin scoring of the same samples (85.2% agreement). Moreover, specific characteristics of each phenotype were identified. While TNBS, oxazolone, and transfer colitis showed high cellularity in stroma, epithelial damage seemed specific for chronic, acute DSS and transfer colitis. Crypt deformations were mostly observed in acute DSS. CONCLUSIONS Quantification of label-free imaging is promising for in vivo endoscopy. In the future, this could be valuable for monitoring of inflammatory pathways in murine models, which is highly relevant for the development of new inflammatory bowel disease therapeutics.
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Affiliation(s)
- Lucas Kreiss
- Address correspondence to: Lucas Kreiss, Dr.-Ing, Institute of Medical Biotechnology, Paul-Gordan-Str 3, 91052 Erlangen, Germany ()
| | | | - Sarah Lemire
- Department of Medicine 1, University Hospital Erlangen, Friedrich-Alexander UniversityErlangen-Nürnberg, Erlangen, Germany,Deutsches Zentrum Immuntherapie, Erlangen, Germany
| | - Kristina Lechner
- Department of Medicine 1, University Hospital Erlangen, Friedrich-Alexander UniversityErlangen-Nürnberg, Erlangen, Germany,Deutsches Zentrum Immuntherapie, Erlangen, Germany
| | - Birgitta Carlé
- Institute of Medical Biotechnology, Friedrich-Alexander UniversityErlangen-Nürnberg, Erlangen, Germany
| | - Ashwathama Dilipkumar
- Institute of Medical Biotechnology, Friedrich-Alexander UniversityErlangen-Nürnberg, Erlangen, Germany,Erlangen Graduate School in Advanced Optical Technologies, Friedrich-Alexander UniversityErlangen-Nürnberg, Erlangen, Germany
| | - Timo Kunert
- Department of Medicine 1, University Hospital Erlangen, Friedrich-Alexander UniversityErlangen-Nürnberg, Erlangen, Germany,Deutsches Zentrum Immuntherapie, Erlangen, Germany
| | - Kristina Scheibe
- Department of Medicine 1, University Hospital Erlangen, Friedrich-Alexander UniversityErlangen-Nürnberg, Erlangen, Germany,Deutsches Zentrum Immuntherapie, Erlangen, Germany
| | - Christina Heichler
- Department of Medicine 1, University Hospital Erlangen, Friedrich-Alexander UniversityErlangen-Nürnberg, Erlangen, Germany,Deutsches Zentrum Immuntherapie, Erlangen, Germany
| | - Anna-Lena Merten
- Institute of Medical Biotechnology, Friedrich-Alexander UniversityErlangen-Nürnberg, Erlangen, Germany,Erlangen Graduate School in Advanced Optical Technologies, Friedrich-Alexander UniversityErlangen-Nürnberg, Erlangen, Germany
| | - Benno Weigmann
- Department of Medicine 1, University Hospital Erlangen, Friedrich-Alexander UniversityErlangen-Nürnberg, Erlangen, Germany,Deutsches Zentrum Immuntherapie, Erlangen, Germany
| | - Clemens Neufert
- Department of Medicine 1, University Hospital Erlangen, Friedrich-Alexander UniversityErlangen-Nürnberg, Erlangen, Germany,Deutsches Zentrum Immuntherapie, Erlangen, Germany
| | - Kai Hildner
- Department of Medicine 1, University Hospital Erlangen, Friedrich-Alexander UniversityErlangen-Nürnberg, Erlangen, Germany,Deutsches Zentrum Immuntherapie, Erlangen, Germany
| | - Michael Vieth
- Institute of Pathology, Klinikum Bayreuth, Bayreuth, Germany,Institute of Pathology, University Hospital Erlangen, Friedrich-Alexander UniversityErlangen-Nürnberg, Erlangen, Germany
| | - Markus F Neurath
- Department of Medicine 1, University Hospital Erlangen, Friedrich-Alexander UniversityErlangen-Nürnberg, Erlangen, Germany,Deutsches Zentrum Immuntherapie, Erlangen, Germany
| | - Oliver Friedrich
- Institute of Medical Biotechnology, Friedrich-Alexander UniversityErlangen-Nürnberg, Erlangen, Germany,Erlangen Graduate School in Advanced Optical Technologies, Friedrich-Alexander UniversityErlangen-Nürnberg, Erlangen, Germany
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15
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Wu W, Liu Q, Brandt C, Tang S. Dual-wavelength multimodal multiphoton microscope with SMA-based depth scanning. BIOMEDICAL OPTICS EXPRESS 2022; 13:2754-2771. [PMID: 35774327 PMCID: PMC9203102 DOI: 10.1364/boe.456390] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Received: 02/16/2022] [Revised: 03/19/2022] [Accepted: 03/31/2022] [Indexed: 06/15/2023]
Abstract
We report on a multimodal multiphoton microscopy (MPM) system with depth scanning. The multimodal capability is realized by an Er-doped femtosecond fiber laser with dual output wavelengths of 1580 nm and 790 nm that are responsible for three-photon and two-photon excitation, respectively. A shape-memory-alloy (SMA) actuated miniaturized objective enables the depth scanning capability. Image stacks combined with two-photon excitation fluorescence (TPEF), second harmonic generation (SHG), and third harmonic generation (THG) signals have been acquired from animal, fungus, and plant tissue samples with a maximum depth range over 200 µm.
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Affiliation(s)
- Wentao Wu
- Department of Electrical and Computer Engineering, University of British Columbia, 5500-2332 Main Mall, Vancouver, BC V6 T 1Z4, Canada
| | - Qihao Liu
- Department of Electrical and Computer Engineering, University of British Columbia, 5500-2332 Main Mall, Vancouver, BC V6 T 1Z4, Canada
| | - Christoph Brandt
- Department of Electrical and Computer Engineering, University of British Columbia, 5500-2332 Main Mall, Vancouver, BC V6 T 1Z4, Canada
| | - Shuo Tang
- Department of Electrical and Computer Engineering, University of British Columbia, 5500-2332 Main Mall, Vancouver, BC V6 T 1Z4, Canada
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16
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Jeon J, Kim H, Jang H, Hwang K, Kim K, Park YG, Jeong KH. Handheld laser scanning microscope catheter for real-time and in vivo confocal microscopy using a high definition high frame rate Lissajous MEMS mirror. BIOMEDICAL OPTICS EXPRESS 2022; 13:1497-1505. [PMID: 35414975 PMCID: PMC8973198 DOI: 10.1364/boe.447558] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Received: 11/03/2021] [Revised: 02/09/2022] [Accepted: 02/09/2022] [Indexed: 06/14/2023]
Abstract
A handheld confocal microscope using a rapid MEMS scanning mirror facilitates real-time optical biopsy for simple cancer diagnosis. Here we report a handheld confocal microscope catheter using high definition and high frame rate (HDHF) Lissajous scanning MEMS mirror. The broad resonant frequency region of the fast axis on the MEMS mirror with a low Q-factor facilitates the flexible selection of scanning frequencies. HDHF Lissajous scanning was achieved by selecting the scanning frequencies with high greatest common divisor (GCD) and high total lobe number. The MEMS mirror was fully packaged into a handheld configuration, which was coupled to a home-built confocal imaging system. The confocal microscope catheter allows fluorescence imaging of in vivo and ex vivo mouse tissues with 30 Hz frame rate and 95.4% fill factor at 256 × 256 pixels image, where the lateral resolution is 4.35 μm and the field-of-view (FOV) is 330 μm × 330 μm. This compact confocal microscope can provide diverse handheld microscopic applications for real-time, on-demand, and in vivo optical biopsy.
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Affiliation(s)
- Jaehun Jeon
- Department of Bio and Brain Engineering, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea
- KAIST Institute for Health Science and Technology (KIHST), KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon, 305-701, Republic of Korea
| | - Hyunwoo Kim
- Department of Bio and Brain Engineering, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea
- KAIST Institute for Health Science and Technology (KIHST), KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon, 305-701, Republic of Korea
| | - Hyunwoo Jang
- Department of Bio and Brain Engineering, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea
| | | | - Kyuyoung Kim
- VPIX Medical, Inc, Deajeon, 34873, Republic of Korea
| | - Young-Gyun Park
- Department of Bio and Brain Engineering, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea
| | - Ki-Hun Jeong
- Department of Bio and Brain Engineering, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea
- KAIST Institute for Health Science and Technology (KIHST), KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon, 305-701, Republic of Korea
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17
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Qualitative Comparison of Image Stitching Algorithms for Multi-Camera Systems in Laparoscopy. J Imaging 2022; 8:jimaging8030052. [PMID: 35324607 PMCID: PMC8951246 DOI: 10.3390/jimaging8030052] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/07/2022] [Revised: 02/04/2022] [Accepted: 02/10/2022] [Indexed: 12/05/2022] Open
Abstract
Multi-camera systems were recently introduced into laparoscopy to increase the narrow field of view of the surgeon. The video streams are stitched together to create a panorama that is easier for the surgeon to comprehend. Multi-camera prototypes for laparoscopy use quite basic algorithms and have only been evaluated on simple laparoscopic scenarios. The more recent state-of-the-art algorithms, mainly designed for the smartphone industry, have not yet been evaluated in laparoscopic conditions. We developed a simulated environment to generate a dataset of multi-view images displaying a wide range of laparoscopic situations, which is adaptable to any multi-camera system. We evaluated classical and state-of-the-art image stitching techniques used in non-medical applications on this dataset, including one unsupervised deep learning approach. We show that classical techniques that use global homography fail to provide a clinically satisfactory rendering and that even the most recent techniques, despite providing high quality panorama images in non-medical situations, may suffer from poor alignment or severe distortions in simulated laparoscopic scenarios. We highlight the main advantages and flaws of each algorithm within a laparoscopic context, identify the main remaining challenges that are specific to laparoscopy, and propose methods to improve these approaches. We provide public access to the simulated environment and dataset.
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18
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Kowalski B, Akondi V, Dubra A. Correction of non-uniform angular velocity and sub-pixel jitter in optical scanning. OPTICS EXPRESS 2022; 30:112-124. [PMID: 35201185 PMCID: PMC8970701 DOI: 10.1364/oe.446162] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/15/2021] [Revised: 12/10/2021] [Accepted: 12/10/2021] [Indexed: 05/08/2023]
Abstract
Optical scanners are widely used in high-resolution scientific, medical, and industrial devices. The accuracy and precision of these instruments are often limited by angular speed fluctuations due to rotational inertia and by poor synchronization between scanning and light detection, respectively. Here we demonstrate that both problems can be mitigated by recording scanner orientation in synchrony with light detection, followed by data resampling. This approach is illustrated with synthetic and experimental data from a point-scanning microscope with a resonant scanner and a non-resonant scanner. Fitting of the resonant scanner orientation data to a cosine model was used to correct image warping and sampling jitter, as well as to precisely interleave image lines collected during the clockwise and counterclockwise resonant scanner portions of the rotation cycle. Vertical scanner orientation data interpolation was used to correct image distortion due to angular speed fluctuations following abrupt control signal changes.
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Affiliation(s)
| | - Vyas Akondi
- Byers Eye Institute, Stanford University, Palo Alto, CA 94303, USA
| | - Alfredo Dubra
- Byers Eye Institute, Stanford University, Palo Alto, CA 94303, USA
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19
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Pshenay-Severin E, Bae H, Reichwald K, Matz G, Bierlich J, Kobelke J, Lorenz A, Schwuchow A, Meyer-Zedler T, Schmitt M, Messerschmidt B, Popp J. Multimodal nonlinear endomicroscopic imaging probe using a double-core double-clad fiber and focus-combining micro-optical concept. LIGHT, SCIENCE & APPLICATIONS 2021; 10:207. [PMID: 34611136 PMCID: PMC8492681 DOI: 10.1038/s41377-021-00648-w] [Citation(s) in RCA: 23] [Impact Index Per Article: 5.8] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 05/12/2021] [Revised: 09/06/2021] [Accepted: 09/15/2021] [Indexed: 05/05/2023]
Abstract
Multimodal non-linear microscopy combining coherent anti-Stokes Raman scattering, second harmonic generation, and two-photon excited fluorescence has proved to be a versatile and powerful tool enabling the label-free investigation of tissue structure, molecular composition, and correlation with function and disease status. For a routine medical application, the implementation of this approach into an in vivo imaging endoscope is required. However, this is a difficult task due to the requirements of a multicolour ultrashort laser delivery from a compact and robust laser source through a fiber with low losses and temporal synchronization, the efficient signal collection in epi-direction, the need for small-diameter but highly corrected endomicroobjectives of high numerical aperture and compact scanners. Here, we introduce an ultra-compact fiber-scanning endoscope platform for multimodal non-linear endomicroscopy in combination with a compact four-wave mixing based fiber laser. The heart of this fiber-scanning endoscope is an in-house custom-designed, single mode, double clad, double core pure silica fiber in combination with a 2.4 mm diameter NIR-dual-waveband corrected endomicroscopic objective of 0.55 numerical aperture and 180 µm field of view for non-linear imaging, allowing a background free, low-loss, high peak power laser delivery, and an efficient signal collection in backward direction. A linear diffractive optical grating overlays pump and Stokes laser foci across the full field of view, such that diffraction-limited performance is demonstrated for tissue imaging at one frame per second with sub-micron spatial resolution and at a high transmission of 65% from the laser to the specimen using a distal resonant fiber scanner.
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Affiliation(s)
| | - Hyeonsoo Bae
- Leibniz Institute of Photonic Technology, Member of Leibniz Health Technologies, Albert-Einstein-Str. 9, 07745, Jena, Germany
| | | | - Gregor Matz
- GRINTECH GmbH, Schillerstr. 1, 07745, Jena, Germany
| | - Jörg Bierlich
- Leibniz Institute of Photonic Technology, Member of Leibniz Health Technologies, Albert-Einstein-Str. 9, 07745, Jena, Germany
| | - Jens Kobelke
- Leibniz Institute of Photonic Technology, Member of Leibniz Health Technologies, Albert-Einstein-Str. 9, 07745, Jena, Germany
| | - Adrian Lorenz
- Leibniz Institute of Photonic Technology, Member of Leibniz Health Technologies, Albert-Einstein-Str. 9, 07745, Jena, Germany
| | - Anka Schwuchow
- Leibniz Institute of Photonic Technology, Member of Leibniz Health Technologies, Albert-Einstein-Str. 9, 07745, Jena, Germany
| | - Tobias Meyer-Zedler
- Leibniz Institute of Photonic Technology, Member of Leibniz Health Technologies, Albert-Einstein-Str. 9, 07745, Jena, Germany
- Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich Schiller University Jena, Helmholtzweg 4, 07743, Jena, Germany
| | - Michael Schmitt
- Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich Schiller University Jena, Helmholtzweg 4, 07743, Jena, Germany
| | | | - Juergen Popp
- Leibniz Institute of Photonic Technology, Member of Leibniz Health Technologies, Albert-Einstein-Str. 9, 07745, Jena, Germany.
- Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich Schiller University Jena, Helmholtzweg 4, 07743, Jena, Germany.
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20
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Wang Y, Li P, Gupta U, Ouyang J, Zhu J. Tunable Soft Lens of Large Focal Length Change. Soft Robot 2021; 9:705-712. [PMID: 34388046 DOI: 10.1089/soro.2021.0036] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022] Open
Abstract
Tunable lens technology inspired by the human eye has opened a new paradigm of smart optical devices for a variety of applications due to unique characteristics such as lightweight, low cost, and facile fabrication over conventional lens assemblies. The fast-growing demands for tunable optical lenses in consumer electronics, medical diagnostics, and optical communications require the lens to have a large focal length modulation range and high compactness. Herein, for the first time, an all-solid tunable soft lens driven by highly transparent dielectric elastomer actuators (DEAs) based on poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) and waterborne polyurethane (PEDOT:PSS/WPU) transparent electrodes is developed. The deformation of the tunable soft lens is achieved by the actuation of DEAs, mimicking the change of the surface profile of the human eye to achieve remarkable focal length variations. Upon electrical activation, this tunable soft lens can vary its original focal length by 209%, which is one of the highest among current tunable soft lenses and far beyond that of the human eye. This study demonstrates that transparent DEAs are capable of achieving focus-variation functions, and potentially useful in artificial robotic vision, visual prostheses, and adjustable glasses, which will induce significant effects on the future development of tunable optics.
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Affiliation(s)
- Yuzhe Wang
- Department of Mechanical Engineering, National University of Singapore, Singapore, Singapore
| | - Pengcheng Li
- Department of Materials Science and Engineering, National University of Singapore, Singapore, Singapore.,Hubei Key Laboratory of Plasma Chemistry and Advanced Materials, School of Materials Science and Engineering, Wuhan Institute of Technology, Wuhan, China
| | - Ujjaval Gupta
- Department of Mechanical Engineering, National University of Singapore, Singapore, Singapore.,Engineering Product Development Pillar, Singapore University of Technology and Design, Singapore, Singapore
| | - Jianyong Ouyang
- Department of Materials Science and Engineering, National University of Singapore, Singapore, Singapore
| | - Jian Zhu
- School of Science and Engineering, Chinese University of Hong Kong at Shenzhen, Shenzhen, China.,Shenzhen Institute of Artificial Intelligence and Robotics for Society, Shenzhen, China
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21
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Li A, Guan H, Park HC, Yue Y, Chen D, Liang W, Li MJ, Lu H, Li X. Twist-free ultralight two-photon fiberscope enabling neuroimaging on freely rotating/walking mice. OPTICA 2021; 8:870-879. [PMID: 39830584 PMCID: PMC11741673 DOI: 10.1364/optica.422657] [Citation(s) in RCA: 15] [Impact Index Per Article: 3.8] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 02/12/2021] [Accepted: 05/12/2021] [Indexed: 01/22/2025]
Abstract
Lightweight and head-mountable scanning nonlinear fiberscope technologies offer an exciting opportunity for enabling mechanistic exploration of ensemble neural activities with subcellular resolution on freely behaving rodents. The tether of the fiberscope, consisting of an optical fiber and scanner drive wires, however, restricts the mouse's movement and consequently precludes free rotation and limits the freedom of walking. Here we present the first twist-free two-photon fiberscope technology for enabling neuroimaging on freely rotating/walking mice. The technology equips a scanning fiberscope with active rotational tracking and compensation capabilities through an optoelectrical commutator (OEC) to allow the animal to rotate and walk in arbitrary patterns during two-photon fluorescence (TPF) imaging of neural activities. The OEC provides excellent optical coupling stability (<±1% fluctuation during rotation) and an extremely high torque sensitivity (<8 mN · m). In addition, the new technology is equipped with a custom grating and prism to effectively manage the temporal properties of the femtosecond excitation pulses through the fiber-optic system, which improved neuroimaging signal by more than 2X. This TPF fiberscope imaging platform has been tested for in vivo imaging, and the results demonstrate that it enables reliable recording of calcium dynamics of more than 50 neurons simultaneously in the motor cortices of freely behaving mice in a twist-free fashion. With active tracking function of the OEC enabled, we observed considerable increase in both behavior and neural activities in the motor cortices of the mice during freely behaving neuroimaging experiments.
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Affiliation(s)
- Ang Li
- Department of Biomedical Engineering, Johns Hopkins University, Baltimore, Maryland 21205, USA
| | - Honghua Guan
- Department of Electrical and Computer Engineering, Johns Hopkins University, Baltimore, Maryland 21218, USA
| | - Hyeon-Cheol Park
- Department of Biomedical Engineering, Johns Hopkins University, Baltimore, Maryland 21205, USA
| | - Yuanlei Yue
- Department of Pharmacology and Physiology, George Washington University, Washington, DC 20052, USA
| | - Defu Chen
- Department of Biomedical Engineering, Johns Hopkins University, Baltimore, Maryland 21205, USA
| | - Wenxuan Liang
- Department of Biomedical Engineering, Johns Hopkins University, Baltimore, Maryland 21205, USA
| | - Ming-Jun Li
- Science and Technology Division, Corning Incorporated, Corning, New York 14831, USA
| | - Hui Lu
- Department of Pharmacology and Physiology, George Washington University, Washington, DC 20052, USA
| | - Xingde Li
- Department of Biomedical Engineering, Johns Hopkins University, Baltimore, Maryland 21205, USA
- Department of Electrical and Computer Engineering, Johns Hopkins University, Baltimore, Maryland 21218, USA
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22
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Kaur M, Lane PM, Menon C. Scanning and Actuation Techniques for Cantilever-Based Fiber Optic Endoscopic Scanners-A Review. SENSORS 2021; 21:s21010251. [PMID: 33401728 PMCID: PMC7795415 DOI: 10.3390/s21010251] [Citation(s) in RCA: 15] [Impact Index Per Article: 3.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 11/23/2020] [Revised: 12/30/2020] [Accepted: 12/30/2020] [Indexed: 01/20/2023]
Abstract
Endoscopes are used routinely in modern medicine for in-vivo imaging of luminal organs. Technical advances in the micro-electro-mechanical system (MEMS) and optical fields have enabled the further miniaturization of endoscopes, resulting in the ability to image previously inaccessible small-caliber luminal organs, enabling the early detection of lesions and other abnormalities in these tissues. The development of scanning fiber endoscopes supports the fabrication of small cantilever-based imaging devices without compromising the image resolution. The size of an endoscope is highly dependent on the actuation and scanning method used to illuminate the target image area. Different actuation methods used in the design of small-sized cantilever-based endoscopes are reviewed in this paper along with their working principles, advantages and disadvantages, generated scanning patterns, and applications.
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Affiliation(s)
- Mandeep Kaur
- MENRVA Research Group, Schools of Mechatronic Systems Engineering and Engineering Science, Simon Fraser University, Surrey, B.C. V3T 0A3, Canada;
- School of Engineering Science, Simon Fraser University, Burnaby, B.C. V5A 1S6, Canada;
- Imaging Unit, Integrative Oncology, BC Cancer Research Center, Vancouver, B.C., V5Z 1L3, Canada
| | - Pierre M. Lane
- School of Engineering Science, Simon Fraser University, Burnaby, B.C. V5A 1S6, Canada;
- Imaging Unit, Integrative Oncology, BC Cancer Research Center, Vancouver, B.C., V5Z 1L3, Canada
| | - Carlo Menon
- MENRVA Research Group, Schools of Mechatronic Systems Engineering and Engineering Science, Simon Fraser University, Surrey, B.C. V3T 0A3, Canada;
- School of Engineering Science, Simon Fraser University, Burnaby, B.C. V5A 1S6, Canada;
- Correspondence:
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23
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Sahu SP, Liu Q, Prasad A, Hasan SMA, Liu Q, Rodriguez MXB, Mukhopadhyay O, Burk D, Francis J, Mukhopadhyay S, Fu X, Gartia MR. Characterization of fibrillar collagen isoforms in infarcted mouse hearts using second harmonic generation imaging. BIOMEDICAL OPTICS EXPRESS 2021; 12:604-618. [PMID: 33520391 PMCID: PMC7818962 DOI: 10.1364/boe.410347] [Citation(s) in RCA: 9] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 09/21/2020] [Revised: 12/14/2020] [Accepted: 12/14/2020] [Indexed: 06/12/2023]
Abstract
We utilized collagen specific second harmonic generation (SHG) signatures coupled with correlative immunofluorescence imaging techniques to characterize collagen structural isoforms (type I and type III) in a murine model of myocardial infarction (MI). Tissue samples were imaged over a four week period using SHG, transmitted light microscopy and immunofluorescence imaging using fluorescently-labeled collagen antibodies. The post-mortem cardiac tissue imaging using SHG demonstrated a progressive increase in collagen deposition in the left ventricle (LV) post-MI. We were able to monitor structural morphology and LV remodeling parameters in terms of extent of LV dilation, stiffness and fiber dimensions in the infarcted myocardium.
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Affiliation(s)
- Sushant P Sahu
- Department of Chemistry, University of Louisiana at Lafayette, Lafayette, LA 70504, USA
| | - Qianglin Liu
- LSU AgCenter, School of Animal Sciences, Louisiana State University, Baton Rouge, LA 70803, USA
| | - Alisha Prasad
- Department of Mechanical and Industrial Engineering, Louisiana State University, Baton Rouge, LA 70803, USA
| | - Syed Mohammad Abid Hasan
- Department of Mechanical and Industrial Engineering, Louisiana State University, Baton Rouge, LA 70803, USA
| | - Qun Liu
- Department of Computer Science, Louisiana State University, Baton Rouge, LA 70803, USA
| | | | | | - David Burk
- Shared Instrumentation Facility and Pennington Biomedical Research Center, Baton Rouge, LA 70808, USA
| | - Joseph Francis
- Comparative Biomedical Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA 70803, USA
| | - Supratik Mukhopadhyay
- Department of Computer Science, Louisiana State University, Baton Rouge, LA 70803, USA
| | - Xing Fu
- LSU AgCenter, School of Animal Sciences, Louisiana State University, Baton Rouge, LA 70803, USA
| | - Manas Ranjan Gartia
- Department of Mechanical and Industrial Engineering, Louisiana State University, Baton Rouge, LA 70803, USA
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24
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Liang W, Park HC, Li K, Li A, Chen D, Guan H, Yue Y, Gau YTA, Bergles DE, Li MJ, Lu H, Li X. Throughput-Speed Product Augmentation for Scanning Fiber-Optic Two-Photon Endomicroscopy. IEEE TRANSACTIONS ON MEDICAL IMAGING 2020; 39:3779-3787. [PMID: 32746124 PMCID: PMC7773217 DOI: 10.1109/tmi.2020.3005067] [Citation(s) in RCA: 14] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/03/2023]
Abstract
Compactness, among several others, is one unique and very attractive feature of a scanning fiber-optic two-photon endomicroscope. To increase the scanning area and the total number of resolvable pixels (i.e., the imaging throughput), it typically requires a longer cantilever which, however, leads to a much undesired, reduced scanning speed (and thus imaging frame rate). Herein we introduce a new design strategy for a fiber-optic scanning endomicroscope, where the overall numerical aperture (NA) or beam focusing power is distributed over two stages: 1) a mode-field focuser engineered at the tip of a double-clad fiber (DCF) cantilever to pre-amplify the single-mode core NA, and 2) a micro objective of a lower magnification (i.e., ∼ 2× in this design) to achieve final tight beam focusing. This new design enables either an ~9-fold increase in imaging area (throughput) or an ~3-fold improvement in imaging frame rate when compared to traditional fiber-optic endomicroscope designs. The performance of an as-designed endomicroscope of an enhanced throughput-speed product was demonstrated by two representative applications: (1) high-resolution imaging of an internal organ (i.e., mouse kidney) in vivo over a large field of view without using any fluorescent contrast agents, and (2) real-time neural imaging by visualizing dendritic calcium dynamics in vivo with sub-second temporal resolution in GCaMP6m-expressing mouse brain. This cascaded NA amplification strategy is universal and can be readily adapted to other types of fiber-optic scanners in compact linear or nonlinear endomicroscopes.
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25
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Pal R, Villarreal P, Yu X, Qiu S, Vargas G. Multimodal widefield fluorescence imaging with nonlinear optical microscopy workflow for noninvasive oral epithelial neoplasia detection: a preclinical study. JOURNAL OF BIOMEDICAL OPTICS 2020; 25:JBO-200213R. [PMID: 33200597 PMCID: PMC7667429 DOI: 10.1117/1.jbo.25.11.116008] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 07/07/2020] [Accepted: 10/02/2020] [Indexed: 05/06/2023]
Abstract
SIGNIFICANCE Early detection of epithelial cancers and precancers/neoplasia in the presence of benign lesions is challenging due to the lack of robust in vivo imaging and biopsy guidance techniques. Label-free nonlinear optical microscopy (NLOM) has shown promise for optical biopsy through the detection of cellular and extracellular signatures of neoplasia. Although in vivo microscopy techniques continue to be developed, the surface area imaged in microscopy is limited by the field of view. FDA-approved widefield fluorescence (WF) imaging systems that capture autofluorescence signatures of neoplasia provide molecular information at large fields of view, which may complement the cytologic and architectural information provided by NLOM. AIM A multimodal imaging approach with high-sensitivity WF and high-resolution NLOM was investigated to identify and distinguish image-based features of neoplasia from normal and benign lesions. APPROACH In vivo label-free WF imaging and NLOM was performed in preclinical hamster models of oral neoplasia and inflammation. Analyses of WF imaging, NLOM imaging, and dual modality (WF combined with NLOM) were performed. RESULTS WF imaging showed increased red-to-green autofluorescence ratio in neoplasia compared to inflammation and normal oral mucosa (p < 0.01). In vivo assessment of the mucosal tissue with NLOM revealed subsurface cytologic (nuclear pleomorphism) and architectural (remodeling of extracellular matrix) atypia in histologically confirmed neoplastic tissue, which were not observed in inflammation or normal mucosa. Univariate and multivariate statistical analysis of macroscopic and microscopic image-based features indicated improved performance (94% sensitivity and 97% specificity) of a multiscale approach over WF alone, even in the presence of benign lesions (inflammation), a common confounding factor in diagnostics. CONCLUSIONS A multimodal imaging approach integrating strengths from WF and NLOM may be beneficial in identifying oral neoplasia. Our study could guide future studies on human oral neoplasia to further evaluate merits and limitations of multimodal workflows and inform the development of multiscale clinical imaging systems.
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Affiliation(s)
- Rahul Pal
- Massachusetts General Hospital and Harvard Medical School, Athinoula A. Martinos Center for Biomedical Imaging, Charlestown, Massachusetts, United States
| | - Paula Villarreal
- The University of Texas Medical Branch, Biomedical Engineering and Imaging Sciences Group, Galveston, Texas, United States
- The University of Texas Medical Branch, Department of Neuroscience, Cell Biology, and Anatomy, Galveston, Texas, United States
| | - Xiaoying Yu
- The University of Texas Medical Branch, Department of Preventive Medicine and Population Health, Galveston, Texas, United States
| | - Suimin Qiu
- The University of Texas Medical Branch, Department of Pathology, Galveston, Texas, United States
| | - Gracie Vargas
- The University of Texas Medical Branch, Biomedical Engineering and Imaging Sciences Group, Galveston, Texas, United States
- The University of Texas Medical Branch, Department of Neuroscience, Cell Biology, and Anatomy, Galveston, Texas, United States
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26
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Endoscopic Optical Imaging Technologies and Devices for Medical Purposes: State of the Art. APPLIED SCIENCES-BASEL 2020. [DOI: 10.3390/app10196865] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/23/2023]
Abstract
The growth and development of optical components and, in particular, the miniaturization of micro-electro-mechanical systems (MEMSs), has motivated and enabled researchers to design smaller and smaller endoscopes. The overarching goal of this work has been to image smaller previously inaccessible luminal organs in real time, at high resolution, in a minimally invasive manner that does not compromise the comfort of the subject, nor introduce additional risk. Thus, an initial diagnosis can be made, or a small precancerous lesion may be detected, in a small-diameter luminal organ that would not have otherwise been possible. Continuous advancement in the field has enabled a wide range of optical scanners. Different scanning techniques, working principles, and the applications of endoscopic scanners are summarized in this review.
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27
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Le Fur M, Zhou IY, Catalano O, Caravan P. Toward Molecular Imaging of Intestinal Pathology. Inflamm Bowel Dis 2020; 26:1470-1484. [PMID: 32793946 PMCID: PMC7500524 DOI: 10.1093/ibd/izaa213] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 04/24/2020] [Indexed: 12/13/2022]
Abstract
Inflammatory bowel disease (IBD) is defined by a chronic relapsing and remitting inflammation of the gastrointestinal tract, with intestinal fibrosis being a major complication. The etiology of IBD remains unknown, but it is thought to arise from a dysregulated and excessive immune response to gut luminal microbes triggered by genetic and environmental factors. To date, IBD has no cure, and treatments are currently directed at relieving symptoms and treating inflammation. The current diagnostic of IBD relies on endoscopy, which is invasive and does not provide information on the presence of extraluminal complications and molecular aspect of the disease. Cross-sectional imaging modalities such as computed tomography enterography (CTE), magnetic resonance enterography (MRE), positron emission tomography (PET), single photon emission computed tomography (SPECT), and hybrid modalities have demonstrated high accuracy for the diagnosis of IBD and can provide both functional and morphological information when combined with the use of molecular imaging probes. This review presents the state-of-the-art imaging techniques and molecular imaging approaches in the field of IBD and points out future directions that could help improve our understanding of IBD pathological processes, along with the development of efficient treatments.
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Affiliation(s)
- Mariane Le Fur
- The Athinoula A. Martinos Center for Biomedical Imaging, Department of Radiology, The Institute for Innovation in Imaging, Massachusetts General Hospital and Harvard Medical School, MA, USA
| | - Iris Y Zhou
- The Athinoula A. Martinos Center for Biomedical Imaging, Department of Radiology, The Institute for Innovation in Imaging, Massachusetts General Hospital and Harvard Medical School, MA, USA
| | - Onofrio Catalano
- The Athinoula A. Martinos Center for Biomedical Imaging, Department of Radiology, The Institute for Innovation in Imaging, Massachusetts General Hospital and Harvard Medical School, MA, USA,The Division of Abdominal Imaging, Department of Radiology, Massachusetts General Hospital and Harvard Medical School, MA, USA
| | - Peter Caravan
- The Athinoula A. Martinos Center for Biomedical Imaging, Department of Radiology, The Institute for Innovation in Imaging, Massachusetts General Hospital and Harvard Medical School, MA, USA,Address correspondence to: Peter Caravan, PhD, The Athinoula A. Martinos Center for Biomedical Imaging, Department of Radiology, The Institute for Innovation in Imaging, Massachusetts General Hospital and Harvard Medical School, 149 Thirteenth Street, Charlestown 02129, MA, USA. E-mail:
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28
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Matsui T, Tamoto R, Iwasa A, Mimura M, Taniguchi S, Hasegawa T, Sudo T, Mizuno H, Kikuta J, Onoyama I, Okugawa K, Shiomi M, Matsuzaki S, Morii E, Kimura T, Kato K, Kiyota Y, Ishii M. Nonlinear Optics with Near-Infrared Excitation Enable Real-Time Quantitative Diagnosis of Human Cervical Cancers. Cancer Res 2020; 80:3745-3754. [PMID: 32718995 DOI: 10.1158/0008-5472.can-20-0348] [Citation(s) in RCA: 11] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/30/2020] [Revised: 05/16/2020] [Accepted: 06/29/2020] [Indexed: 11/16/2022]
Abstract
Histopathologic analysis through biopsy has been one of the most useful methods for the assessment of malignant neoplasms. However, some aspects of the analysis such as invasiveness, evaluation range, and turnaround time from biopsy to report could be improved. Here, we report a novel method for visualizing human cervical tissue three-dimensionally, without biopsy, fixation, or staining, and with sufficient quality for histologic diagnosis. Near-infrared excitation and nonlinear optics were employed to visualize unstained human epithelial tissues of the cervix uteri by constructing images with third-harmonic generation (THG) and second-harmonic generation (SHG). THG images enabled evaluation of nuclear morphology in a quantitative manner with six parameters after image analysis using deep learning. It was also possible to quantitatively assess intraepithelial fibrotic changes based on SHG images and another deep learning analysis. Using each analytical procedure alone, normal and cancerous tissue were classified quantitatively with an AUC ≥0.92. Moreover, a combinatory analysis of THG and SHG images with a machine learning algorithm allowed accurate classification of three-dimensional image files of normal tissue, intraepithelial neoplasia, and invasive carcinoma with a weighted kappa coefficient of 0.86. Our method enables real-time noninvasive diagnosis of cervical lesions, thus constituting a potential tool to dramatically change early detection. SIGNIFICANCE: This study proposes a novel method for diagnosing cancer using nonlinear optics, which enables visualization of histologic features of living tissues without the need for any biopsy or staining dye.
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Affiliation(s)
- Takahiro Matsui
- Department of Immunology and Cell Biology, Osaka University Graduate School of Medicine, Suita, Osaka, Japan
| | - Ryo Tamoto
- Yokohama Plant, Nikon Corporation, Yokohama, Kanagawa, Japan
| | - Akio Iwasa
- Yokohama Plant, Nikon Corporation, Yokohama, Kanagawa, Japan
| | - Masafumi Mimura
- Yokohama Plant, Nikon Corporation, Yokohama, Kanagawa, Japan
| | - Seiji Taniguchi
- Department of Immunology and Cell Biology, Osaka University Graduate School of Medicine, Suita, Osaka, Japan
| | - Tetsuo Hasegawa
- Department of Immunology and Cell Biology, Osaka University Graduate School of Medicine, Suita, Osaka, Japan
| | - Takao Sudo
- Department of Immunology and Cell Biology, Osaka University Graduate School of Medicine, Suita, Osaka, Japan
| | - Hiroki Mizuno
- Department of Immunology and Cell Biology, Osaka University Graduate School of Medicine, Suita, Osaka, Japan
| | - Junichi Kikuta
- Department of Immunology and Cell Biology, Osaka University Graduate School of Medicine, Suita, Osaka, Japan
| | - Ichiro Onoyama
- Department of Obstetrics and Gynecology, Kyushu University Graduate School of Medical Sciences, Higashi-ku, Fukuoka, Japan
| | - Kaoru Okugawa
- Department of Obstetrics and Gynecology, Kyushu University Graduate School of Medical Sciences, Higashi-ku, Fukuoka, Japan
| | - Mayu Shiomi
- Department of Obstetrics and Gynecology, Osaka University Graduate School of Medicine, Suita, Osaka, Japan
| | - Shinya Matsuzaki
- Department of Obstetrics and Gynecology, Osaka University Graduate School of Medicine, Suita, Osaka, Japan
| | - Eiichi Morii
- Department of Pathology, Osaka University Graduate School of Medicine, Suita, Osaka, Japan
| | - Tadashi Kimura
- Department of Obstetrics and Gynecology, Osaka University Graduate School of Medicine, Suita, Osaka, Japan
| | - Kiyoko Kato
- Department of Obstetrics and Gynecology, Kyushu University Graduate School of Medical Sciences, Higashi-ku, Fukuoka, Japan
| | - Yasujiro Kiyota
- Yokohama Plant, Nikon Corporation, Yokohama, Kanagawa, Japan
| | - Masaru Ishii
- Department of Immunology and Cell Biology, Osaka University Graduate School of Medicine, Suita, Osaka, Japan.
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29
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Park HC, Guan H, Li A, Yue Y, Li MJ, Lu H, Li X. High-speed fiber-optic scanning nonlinear endomicroscopy for imaging neuron dynamicsin vivo. OPTICS LETTERS 2020; 45:3605-3608. [PMID: 32630910 PMCID: PMC7585368 DOI: 10.1364/ol.396023] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 04/24/2020] [Accepted: 05/23/2020] [Indexed: 06/11/2023]
Abstract
Fiber-optic-based two-photon fluorescence endomicroscopy is emerging as an enabling technology for in vivo histological imaging of internal organs and functional neuronal imaging on freely-behaving animals. However, high-speed imaging remains challenging due to the expense of miniaturization and lack of suited fast beam scanners. For many applications, a higher imaging speed is highly desired, especially for monitoring functional dynamics such as transient dendritic responses in neuroscience. This Letter reports the development of a fast fiber-optic scanning endo-microscope with an imaging speed higher than 26 frames/s. In vivo neural dynamics imaging with the high-speed endomicroscope was performed on a freely-behaving mouse over the primary motor cortex that expressed GCaMP6m. The results demonstrate its capability of real-time monitoring of transient neuronal dynamics with very fine temporal resolution.
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Affiliation(s)
- Hyeon-Cheol Park
- Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA
| | - Honghua Guan
- Department of Electrical and Computer Engineering, Johns Hopkins University, Baltimore, Maryland 21218, USA
| | - Ang Li
- Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA
| | - Yuanlei Yue
- Department of Pharmacology and Physiology, George Washington University School of Medicine, Washington, DC 20052, USA
| | - Ming-Jun Li
- Science and Technology Division, Corning Incorporated, Corning, New York 14831, USA
| | - Hui Lu
- Department of Pharmacology and Physiology, George Washington University School of Medicine, Washington, DC 20052, USA
| | - Xingde Li
- Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA
- Department of Electrical and Computer Engineering, Johns Hopkins University, Baltimore, Maryland 21218, USA
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30
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Kudlinski A, Cassez A, Vanvincq O, Septier D, Pastre A, Habert R, Baudelle K, Douay M, Mytskaniuk V, Tsvirkun V, Rigneault H, Bouwmans G. Double clad tubular anti-resonant hollow core fiber for nonlinear microendoscopy. OPTICS EXPRESS 2020; 28:15062-15070. [PMID: 32403539 DOI: 10.1364/oe.389084] [Citation(s) in RCA: 15] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/12/2023]
Abstract
We report the fabrication and characterization of the first double clad tubular anti-resonant hollow core fiber. It allows to deliver ultrashort pulses without temporal nor spectral distortions in the 700-1000 nm wavelength range and to efficiently collect scattered light in a high numerical aperture double clad. The output fiber mode is shaped with a silica microsphere generating a photonic nanojet, making it well suitable for nonlinear microendoscopy application. Additionally, we provide an open access software allowing to find optimal drawing parameters for the fabrication of tubular hollow core fibers.
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31
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Pouli D, Thieu HT, Genega EM, Baecher-Lind L, House M, Bond B, Roncari DM, Evans ML, Rius-Diaz F, Munger K, Georgakoudi I. Label-free, High-Resolution Optical Metabolic Imaging of Human Cervical Precancers Reveals Potential for Intraepithelial Neoplasia Diagnosis. CELL REPORTS MEDICINE 2020; 1. [PMID: 32577625 PMCID: PMC7311071 DOI: 10.1016/j.xcrm.2020.100017] [Citation(s) in RCA: 14] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Indexed: 01/07/2023]
Abstract
While metabolic changes are considered a cancer hallmark, their assessment has not been incorporated in the detection of early or precancers, when treatment is most effective. Here, we demonstrate that metabolic changes are detected in freshly excised human cervical precancerous tissues using label-free, non-destructive imaging of the entire epithelium. The images rely on two-photon excited fluorescence from two metabolic co-enzymes, NAD(P)H and FAD, and have micron-level resolution, enabling sensitive assessments of the redox ratio and mitochondrial fragmentation, which yield metrics of metabolic function and heterogeneity. Simultaneous characterization of morphological features, such as the depth-dependent variation of the nuclear:cytoplasmic ratio, is demonstrated. Multi-parametric analysis combining several metabolic metrics with morphological ones enhances significantly the diagnostic accuracy of identifying high-grade squamous intraepithelial lesions. Our results motivate the translation of such functional metabolic imaging to in vivo studies, which may enable improved identification of cervical lesions, and other precancers, at the bedside.
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Affiliation(s)
- Dimitra Pouli
- Department of Biomedical Engineering, Tufts University, 4 Colby Street, Medford, MA 02155, USA.,Present address: Department of Pathology, Beth Israel Deaconess Medical Center and Harvard Medical School, 330 Brookline Avenue, Boston, MA 02115, USA
| | - Hong-Thao Thieu
- Department of Obstetrics and Gynecology, Tufts University School of Medicine, Tufts Medical Center, 800 Washington Street, Boston, MA 02111, USA
| | - Elizabeth M Genega
- Department of Pathology and Laboratory Medicine, Tufts University School of Medicine, Tufts Medical Center, 800 Washington Street, Boston, MA 02111, USA
| | - Laura Baecher-Lind
- Department of Obstetrics and Gynecology, Tufts University School of Medicine, Tufts Medical Center, 800 Washington Street, Boston, MA 02111, USA
| | - Michael House
- Department of Obstetrics and Gynecology, Tufts University School of Medicine, Tufts Medical Center, 800 Washington Street, Boston, MA 02111, USA
| | - Brian Bond
- Department of Obstetrics and Gynecology, Tufts University School of Medicine, Tufts Medical Center, 800 Washington Street, Boston, MA 02111, USA.,Present address: Department of Obstetrics and Gynecology, University of Massachusetts School of Medicine, 55 Lake Avenue North, Worcester, MA 01655, USA
| | - Danielle M Roncari
- Department of Obstetrics and Gynecology, Tufts University School of Medicine, Tufts Medical Center, 800 Washington Street, Boston, MA 02111, USA
| | - Megan L Evans
- Department of Obstetrics and Gynecology, Tufts University School of Medicine, Tufts Medical Center, 800 Washington Street, Boston, MA 02111, USA
| | - Francisca Rius-Diaz
- Department of Preventive Medicine and Public Health, Faculty of Medicine, University of Málaga, 32 Louis Pasteur Boulevard, 29071 Málaga, Spain
| | - Karl Munger
- Department of Developmental, Molecular, and Chemical Biology, Tufts University School of Medicine, 136 Harrison Avenue, Boston, MA 02111, USA
| | - Irene Georgakoudi
- Department of Biomedical Engineering, Tufts University, 4 Colby Street, Medford, MA 02155, USA.,Lead Contact
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32
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Glover B, Teare J, Patel N. The Status of Advanced Imaging Techniques for Optical Biopsy of Colonic Polyps. Clin Transl Gastroenterol 2020; 11:e00130. [PMID: 32352708 PMCID: PMC7145035 DOI: 10.14309/ctg.0000000000000130] [Citation(s) in RCA: 15] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 08/16/2019] [Accepted: 01/03/2020] [Indexed: 12/19/2022] Open
Abstract
The progressive miniaturization of photonic components presents the opportunity to obtain unprecedented microscopic images of colonic polyps in real time during endoscopy. This information has the potential to act as "optical biopsy" to aid clinical decision-making, including the possibility of adopting new paradigms such as a "resect and discard" approach for low-risk lesions. The technologies discussed in this review include confocal laser endomicroscopy, optical coherence tomography, multiphoton microscopy, Raman spectroscopy, and hyperspectral imaging. These are in different stages of development and clinical readiness, but all show the potential to produce reliable in vivo discrimination of different tissue types. A structured literature search of the imaging techniques for colorectal polyps has been conducted. The significant developments in endoscopic imaging were identified for each modality, and the status of current development was discussed. Of the advanced imaging techniques discussed, confocal laser endomicroscopy is in clinical use and, under optimal conditions with an experienced operator, can provide accurate histological assessment of tissue. The remaining techniques show potential for incorporation into endoscopic equipment and practice, although further component development is needed, followed by robust prospective validation of accuracy. Optical coherence tomography illustrates tissue "texture" well and gives good assessment of mucosal thickness and layers. Multiphoton microscopy produces high-resolution images at a subcellular resolution. Raman spectroscopy and hyperspectral imaging are less developed endoscopically but provide a tissue "fingerprint" which can distinguish between tissue types. Molecular imaging may become a powerful adjunct to other techniques, with its ability to precisely label specific molecules within tissue and thereby enhance imaging.
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Affiliation(s)
- Ben Glover
- Department of Surgery and Cancer, Imperial College London, London, UK
| | - Julian Teare
- Department of Surgery and Cancer, Imperial College London, London, UK
| | - Nisha Patel
- Department of Surgery and Cancer, Imperial College London, London, UK
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33
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Zhou G, Lim ZH, Qi Y, Zhou G. Single-Pixel MEMS Imaging Systems. MICROMACHINES 2020; 11:E219. [PMID: 32093324 PMCID: PMC7074650 DOI: 10.3390/mi11020219] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 01/31/2020] [Revised: 02/18/2020] [Accepted: 02/19/2020] [Indexed: 11/16/2022]
Abstract
Single-pixel imaging technology is an attractive technology considering the increasing demand of imagers that can operate in wavelengths where traditional cameras have limited efficiency. Meanwhile, the miniaturization of imaging systems is also desired to build affordable and portable devices for field applications. Therefore, single-pixel imaging systems based on microelectromechanical systems (MEMS) is an effective solution to develop truly miniaturized imagers, owing to their ability to integrate multiple functionalities within a small device. MEMS-based single-pixel imaging systems have mainly been explored in two research directions, namely the encoding-based approach and the scanning-based approach. The scanning method utilizes a variety of MEMS scanners to scan the target scenery and has potential applications in the biological imaging field. The encoding-based system typically employs MEMS modulators and a single-pixel detector to encode the light intensities of the scenery, and the images are constructed by harvesting the power of computational technology. This has the capability to capture non-visible images and 3D images. Thus, this review discusses the two approaches in detail, and their applications are also reviewed to evaluate the efficiency and advantages in various fields.
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Affiliation(s)
- Guangcan Zhou
- Micro and Nano Systems Initiative, Department of Mechanical Engineering, National University of Singapore, Singapore 117575, Singapore; (G.Z.); (Z.H.L.); (Y.Q.)
| | - Zi Heng Lim
- Micro and Nano Systems Initiative, Department of Mechanical Engineering, National University of Singapore, Singapore 117575, Singapore; (G.Z.); (Z.H.L.); (Y.Q.)
| | - Yi Qi
- Micro and Nano Systems Initiative, Department of Mechanical Engineering, National University of Singapore, Singapore 117575, Singapore; (G.Z.); (Z.H.L.); (Y.Q.)
| | - Guangya Zhou
- Micro and Nano Systems Initiative, Department of Mechanical Engineering, National University of Singapore, Singapore 117575, Singapore; (G.Z.); (Z.H.L.); (Y.Q.)
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34
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Zhao Y, Maguluri G, Ferguson RD, Tu H, Paul K, Boppart SA, Llano DA, Iftimia N. Two-photon microscope using a fiber-based approach for supercontinuum generation and light delivery to a small-footprint optical head. OPTICS LETTERS 2020; 45:909-912. [PMID: 32058502 PMCID: PMC7316260 DOI: 10.1364/ol.381571] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/28/2019] [Accepted: 01/06/2020] [Indexed: 05/20/2023]
Abstract
In this Letter, we report a low-cost, portable, two-photon excitation fluorescence microscopy imager that uses a fiber-based approach for both femtosecond supercontinuum (SC) generation and light delivery to the optical head. The SC generation is based on a tapered polarization-maintaining photonic crystal fiber that uses pre-chirped femtosecond narrowband pulses to generate a coherent SC spectrum with a bandwidth of approximately 300 nm. Using this approach, high-power, near-transform-limited, wavelength-selectable SC pulses are generated and directly delivered to the imaging optical head. Preliminary testing of this imager on brain slices is presented, demonstrating a high signal-to-noise ratio and sub-cellular imaging capabilities to a depth of approximately 200 µm. These results demonstrate the suitability of the technology for ex vivo and potentially in vivo cellular-level biomedical imaging applications.
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Affiliation(s)
- Youbo Zhao
- Physical Sciences Inc., 20 New England Business Center Dr., Andover, Massachusetts 01810, USA
| | - Gopi Maguluri
- Physical Sciences Inc., 20 New England Business Center Dr., Andover, Massachusetts 01810, USA
| | - R. Daniel Ferguson
- Physical Sciences Inc., 20 New England Business Center Dr., Andover, Massachusetts 01810, USA
| | - Haohua Tu
- Beckman Institute for Advanced Science and Technology, University of Illinois, 405 N. Mathews Ave., Urbana, Illinois 61822, USA
| | - Kush Paul
- Beckman Institute for Advanced Science and Technology, University of Illinois, 405 N. Mathews Ave., Urbana, Illinois 61822, USA
- Department of Molecular and Integrative Physiology, University of Illinois, 405 N. Mathews Ave., Urbana, Illinois 61822, USA
| | - Stephen A. Boppart
- Beckman Institute for Advanced Science and Technology, University of Illinois, 405 N. Mathews Ave., Urbana, Illinois 61822, USA
| | - Daniel A. Llano
- Beckman Institute for Advanced Science and Technology, University of Illinois, 405 N. Mathews Ave., Urbana, Illinois 61822, USA
- Department of Molecular and Integrative Physiology, University of Illinois, 405 N. Mathews Ave., Urbana, Illinois 61822, USA
| | - Nicusor Iftimia
- Physical Sciences Inc., 20 New England Business Center Dr., Andover, Massachusetts 01810, USA
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35
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He K, Zhao L, Chen Y, Huang X, Ding Y, Hua H, Liu L, Wang X, Wang M, Zhang Y, Fan Z. Label-free multiphoton microscopic imaging as a novel real-time approach for discriminating colorectal lesions: A preliminary study. J Gastroenterol Hepatol 2019; 34:2144-2151. [PMID: 31265738 DOI: 10.1111/jgh.14772] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 10/14/2018] [Revised: 05/21/2019] [Accepted: 06/25/2019] [Indexed: 12/14/2022]
Abstract
BACKGROUND AND AIM Although various endoscopic technologies have been developed to increase the visual diagnostic accuracy of colorectal precancerosis and early carcinoma, the biopsy-dependent pathology still remains the golden standard. During routine endoscopy, real-time optical histological diagnosis is desired. Without fluorescent label, multiphoton microscopy (MPM) imaging directly reveals live cellular morphology and tissue microenvironment based on intrinsic two-photon excited fluorescence and second harmonic generation signals. Its high-imaging resolution and performance are comparable with the histopathology. We thus aimed to initially investigate the original features of colorectal diseases under MPM and evaluate its potential for real-time diagnosis. METHODS Experimental and diagnostic cohorts were designed. Multiphoton images of 40 ex vivo fresh tissues confirmed pathologically of colorectal normal tissues, hyperplastic polyps, adenomas, and adenocarcinomas were collected. Features were recorded to establish diagnostic standards with MPM. For the second cohort with 92 fresh tissues, we distinguished the various colorectal diseases with conclusive MPM features. RESULTS Through the investigation, the colorectal diseases were presented differences in the crypt opening, gland structure, epithelial cells, and collagen fibers. With the typical features, we preliminarily tested the diagnostic efficiency and found that its sensitivity for distinguishing normal, hyperplastic polyps, adenoma and adenocarcinoma was 88.89%, 76.47%, 83.33%, and 97.92%, while the specificity was 99.32%, 94.00%, 94.92%, and 94.12%, respectively. CONCLUSION The real-time multiphoton microscopic imaging can be effective to identify the colorectal lesions with high resolution. Via integrating with the endoscopes in the future, it could promote precise optical diagnosis in clinics.
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Affiliation(s)
- Kexin He
- Department of Digestive Endoscopy, The First Affiliated Hospital with Nanjing Medical University, Nanjing, China
| | - Lili Zhao
- Department of Digestive Endoscopy, The First Affiliated Hospital with Nanjing Medical University, Nanjing, China
| | - Yingtong Chen
- Department of Digestive Endoscopy, The First Affiliated Hospital with Nanjing Medical University, Nanjing, China
| | - Xiaoyang Huang
- National Laboratory of Solid State Microstructure of Nanjing University, Nanjing, China
| | - Ying Ding
- Department of Pathology, The First Affiliated Hospital with Nanjing Medical University, Nanjing, China
| | - Hongjin Hua
- Department of Pathology, The First Affiliated Hospital with Nanjing Medical University, Nanjing, China
| | - Li Liu
- Department of Digestive Endoscopy, The First Affiliated Hospital with Nanjing Medical University, Nanjing, China
| | - Xiang Wang
- Department of Digestive Endoscopy, The First Affiliated Hospital with Nanjing Medical University, Nanjing, China
| | - Min Wang
- Department of Digestive Endoscopy, The First Affiliated Hospital with Nanjing Medical University, Nanjing, China
| | - Yong Zhang
- National Laboratory of Solid State Microstructure of Nanjing University, Nanjing, China
| | - Zhining Fan
- Department of Digestive Endoscopy, The First Affiliated Hospital with Nanjing Medical University, Nanjing, China.,Department of General Surgery, The First Affiliated Hospital with Nanjing Medical University, Nanjing, China
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Sibai M, Mehidine H, Moawad EK, Juchaux M, Varlet P, Devaux B, Abi Haidar D. Comparison of optically-derived biomarkers in healthy and brain tumor tissue under one- and two-photon excitation. JOURNAL OF BIOPHOTONICS 2019; 12:e201900111. [PMID: 31276313 DOI: 10.1002/jbio.201900111] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 03/25/2019] [Revised: 07/02/2019] [Accepted: 07/04/2019] [Indexed: 06/09/2023]
Abstract
The surgical outcome of brain tumor resection and needle biopsy is significantly correlated to the patient's prognosis. Brain tumor surgery is limited to resecting the solid portion of the tumor as current intraoperative imaging modalities are incapable of delineating infiltrative regions. For accurate delineation, in situ tissue interrogation at the submicron scale is warranted. Additionally, multimodal detection is required to remediate the genetically and molecularly heterogeneous nature of brain tumors, notably, that of gliomas, meningioma and brain metastasis. Multimodal detection, such as spectrally- and temporally-resolved fluorescence under one- and two-photon excitation, enables characterizing tissue based on several endogenous optical contrasts. In order to assign the optically-derived parameters to different tissue types, construction of an optical database obtained from biopsied tissue is warranted. This report showcases the different quantitative and semi-quantitative optical markers that may comprise the tissue discrimination database. These include: the optical index ratio, the optical redox ratio, the relative collagen density, spectrally-resolved fluorescence lifetime parameters, two-photon fluorescence imaging and second harmonic generation imaging.
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Affiliation(s)
- Mira Sibai
- Imagerie et Modélisation en Neurobiologie et Cancérologie (IMNC), CNRS, Univ Paris Sud, Université Paris-Saclay, Orsay, France
- Université de Paris, IMNC, Orsay, France
| | - Hussein Mehidine
- Imagerie et Modélisation en Neurobiologie et Cancérologie (IMNC), CNRS, Univ Paris Sud, Université Paris-Saclay, Orsay, France
- Université de Paris, IMNC, Orsay, France
| | - Emile Kaadou Moawad
- Imagerie et Modélisation en Neurobiologie et Cancérologie (IMNC), CNRS, Univ Paris Sud, Université Paris-Saclay, Orsay, France
- Université de Paris, IMNC, Orsay, France
| | - Marjorie Juchaux
- Imagerie et Modélisation en Neurobiologie et Cancérologie (IMNC), CNRS, Univ Paris Sud, Université Paris-Saclay, Orsay, France
- Université de Paris, IMNC, Orsay, France
| | - Pascale Varlet
- Neuropathology Department, Sainte-Anne Hospital, Paris, France
- IMA BRAIN, INSERMU894, Centre de Psychiatrie et de Neurosciences, Paris, France
- Paris Descartes University, Paris, France
| | - Bertrand Devaux
- Paris Descartes University, Paris, France
- Neurosurgery Department, Sainte-Anne Hospital, Paris, France
| | - Darine Abi Haidar
- Imagerie et Modélisation en Neurobiologie et Cancérologie (IMNC), CNRS, Univ Paris Sud, Université Paris-Saclay, Orsay, France
- Université de Paris, IMNC, Orsay, France
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Abstract
BACKGROUND AND AIMS Endoscopic imaging is a rapidly progressing field and benefits from miniaturization of advanced imaging technologies, which may allow accurate real-time characterization of lesions. The concept of the "optical biopsy" to predict polyp histology has gained prominence in recent years and may become clinically applicable with the advent of new imaging technology. This review aims to discuss current evidence and examine the emerging technologies as applied to the optical diagnosis of colorectal polyps. METHODS A structured literature search and review has been carried out of the evidence for diagnostic accuracy of image-enhanced endoscopy and emerging endoscopic imaging technologies. The image-enhanced endoscopy techniques are reviewed, including their basic scientific principles and current evidence for effectiveness. These include the established image-enhancement technologies such as narrow-band imaging, i-scan, and Fuji intelligent chromoendoscopy. More recent technologies including optical enhancement, blue laser imaging, and linked color imaging are discussed. Adjunctive imaging techniques in current clinical use are discussed, such as autofluorescence imaging and endocytoscopy. The emerging advanced imaging techniques are reviewed, including confocal laser endomicroscopy, optical coherence tomography, and Raman spectroscopy. CONCLUSIONS Large studies of the established image-enhancement techniques show some role for the optical diagnosis of polyp histology, although results have been mixed, and at present only the technique of narrow-band imaging is appropriate for the diagnosis of low-risk polyps when used by an expert operator. Other image-enhancement techniques will require further study to validate their accuracy but show potential to support the use of a "resect-and-discard" approach to low-risk polyps. New technologies show exciting potential for real-time diagnosis, but further clinical studies in humans have yet to be performed.
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Andreana M, Sentosa R, Erkkilä MT, Drexler W, Unterhuber A. Depth resolved label-free multimodal optical imaging platform to study morpho-molecular composition of tissue. Photochem Photobiol Sci 2019; 18:997-1008. [PMID: 30882117 DOI: 10.1039/c8pp00410b] [Citation(s) in RCA: 13] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/21/2022]
Abstract
Multimodal imaging platforms offer a vast array of tissue information in a single image acquisition by combining complementary imaging techniques. By merging different systems, better tissue characterization can be achieved than is possible by the constituent imaging modalities alone. The combination of optical coherence tomography (OCT) with non-linear optical imaging (NLOI) techniques such as two-photon excited fluorescence (TPEF), second harmonic generation (SHG) and coherent anti-Stokes Raman scattering (CARS) provides access to detailed information of tissue structure and molecular composition in a fast, label-free and non-invasive manner. We introduce a multimodal label-free approach for morpho-molecular imaging and spectroscopy and validate the system in mouse skin demonstrating the potential of the system for colocalized acquisition of OCT and NLOI signals.
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Affiliation(s)
- Marco Andreana
- Medical University of Vienna, Center for Medical Physics and Biomedical Engineering, Waehringer Guertel 18-20, 1090 Vienna, Austria.
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Prieto SP, Reed CL, James HM, Quinn KP, Muldoon TJ. Differences in colonic crypt morphology of spontaneous and colitis-associated murine models via second harmonic generation imaging to quantify colon cancer development. BMC Cancer 2019; 19:428. [PMID: 31072353 PMCID: PMC6507102 DOI: 10.1186/s12885-019-5639-8] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/14/2018] [Accepted: 04/24/2019] [Indexed: 12/23/2022] Open
Abstract
Background Colorectal cancer remains the second leading cause of cancer death in the United States, and increased risk in patients with ulcerative colitis (a subset of inflammatory bowel disease) has motivated studies into early markers of dysplasia. The development of clinically translatable multiphoton imaging systems has allowed for the potential of in vivo label-free imaging of epithelial crypt structures via autofluorescence and/or second harmonic generation (SHG). SHG has been used to investigate collagen structures in various types of cancer, though the changes that colorectal epithelial collagen structures undergo during tumor development, specifically colitis-associated tumors, have not been fully investigated. Methods This study used two murine models, using A/J mice, one for spontaneous carcinoma and one for colitis-associated carcinoma, to investigate and quantify SHG image features that could potentially inform future study designs of endoscopic multiphoton imaging systems. The spontaneous tumor model comprised a series of six weekly injections of azoxymethane (AOM model). The colitis-associated tumor model comprised a single injection of AOM, followed by cycles of drinking water with dissolved dextran sodium sulfate salt (AOM-DSS model). SHG images of freshly resected murine colon were acquired with a multiphoton imaging system, and image features, such as crypt size, shape and distribution, were quantified using an automated algorithm. Results The comparison of quantified features of crypt morphology demonstrated the ability of our quantitative image feature algorithms to detect differences between spontaneous (AOM model) and colitis-associated (AOM-DSS model) murine colorectal tissue specimens. There were statistically significant differences in the mean and standard deviation of nearest neighbor (distance between crypts) and circularity between the Control cohort, AOM and AOM-DSS cohorts. We also saw significance between AOM and AOM-DSS cohorts when calculating nearest neighbor in images acquired at fixed depths. Conclusion The results provide insight into the ability of SHG imaging to yield relevant data about the crypt microstructure in colorectal epithelium, specifically the potential to distinguish between spontaneous and colitis-associated murine models using quantification of crypt shape and distribution, informing future design of translational multiphoton imaging systems and protocols. Electronic supplementary material The online version of this article (10.1186/s12885-019-5639-8) contains supplementary material, which is available to authorized users.
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Affiliation(s)
- Sandra P Prieto
- Biomedical Engineering Department, University of Arkansas, Fayetteville, AR, 72701, USA
| | - Cassandra L Reed
- Biomedical Engineering Department, University of Arkansas, Fayetteville, AR, 72701, USA
| | - Haley M James
- Biomedical Engineering Department, University of Arkansas, Fayetteville, AR, 72701, USA
| | - Kyle P Quinn
- Biomedical Engineering Department, University of Arkansas, Fayetteville, AR, 72701, USA
| | - Timothy J Muldoon
- Biomedical Engineering Department, University of Arkansas, Fayetteville, AR, 72701, USA.
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40
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Park HC, Zhang X, Yuan W, Zhou L, Xie H, Li X. Ultralow-voltage electrothermal MEMS based fiber-optic scanning probe for forward-viewing endoscopic OCT. OPTICS LETTERS 2019; 44:2232-2235. [PMID: 31042191 PMCID: PMC6541216 DOI: 10.1364/ol.44.002232] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 01/29/2019] [Accepted: 04/01/2019] [Indexed: 06/09/2023]
Abstract
We report an ultralow-voltage, electrothermal (ET) micro-electro-mechanical system (MEMS) based probe for forward-viewing endoscopic optical coherence tomography (OCT) imaging. The fully assembled probe has a diameter of 5.5 mm and a length of 55 mm, including the imaging optics and a 40 mm long fiber-optic cantilever attached on a micro-platform of the bimorph ET MEMS actuator. The ET MEMS actuator provides a sufficient mechanical actuation force as well as a large vertical displacement, achieving up to a 3 mm optical scanning range with only a 3 VACp-p drive voltage with a 1.5 VDC offset. The imaging probe was integrated with a swept-source OCT system of a 100 kHz A-scan rate, and its performance was successfully demonstrated with cross-sectional imaging of biological tissues ex vivo and in vivo at a speed up to 200 frames per second.
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Affiliation(s)
- Hyeon-Cheol Park
- Department of Biomedical Engineering, School of Medicine, Johns Hopkins University, Baltimore, Maryland 21205, USA
| | - Xiaoyang Zhang
- Department of Electrical and Computer Engineering, University of Florida, Gainesville, Florida 32611, USA
| | - Wu Yuan
- Department of Biomedical Engineering, School of Medicine, Johns Hopkins University, Baltimore, Maryland 21205, USA
| | - Liang Zhou
- Department of Electrical and Computer Engineering, University of Florida, Gainesville, Florida 32611, USA
| | - Huikai Xie
- Department of Electrical and Computer Engineering, University of Florida, Gainesville, Florida 32611, USA
| | - Xingde Li
- Department of Biomedical Engineering, School of Medicine, Johns Hopkins University, Baltimore, Maryland 21205, USA
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Dilipkumar A, Al‐Shemmary A, Kreiß L, Cvecek K, Carlé B, Knieling F, Gonzales Menezes J, Thoma O, Schmidt M, Neurath MF, Waldner M, Friedrich O, Schürmann S. Label-Free Multiphoton Endomicroscopy for Minimally Invasive In Vivo Imaging. ADVANCED SCIENCE (WEINHEIM, BADEN-WURTTEMBERG, GERMANY) 2019; 6:1801735. [PMID: 31016109 PMCID: PMC6468963 DOI: 10.1002/advs.201801735] [Citation(s) in RCA: 45] [Impact Index Per Article: 7.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/04/2018] [Revised: 01/29/2019] [Indexed: 05/24/2023]
Abstract
Multiphoton microscopy of cellular autofluorescence and second harmonic generation from collagen facilitates imaging of living cells and tissues without the need for additional fluorescent labels. Here, a compact multiphoton endomicroscope for label-free in vivo imaging in small animals via side-viewing needle objectives is presented. Minimal invasive imaging at cellular resolution is performed in colonoscopy of mice without surgical measures and without fluorescent dyes as a contrast agent. The colon mucosa is imaged repeatedly in the same animal in a mouse model of acute intestinal inflammation to study the process of inflammation at the tissue level within a time period of ten days, demonstrating the capabilities of label-free endomicroscopy for longitudinal studies for the first time.
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Affiliation(s)
- Ashwathama Dilipkumar
- Institute of Medical BiotechnologyFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 391052ErlangenGermany
- Erlangen Graduate School in Advanced Optical TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 791052ErlangenGermany
| | - Alaa Al‐Shemmary
- Institute of Medical BiotechnologyFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 391052ErlangenGermany
- Erlangen Graduate School in Advanced Optical TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 791052ErlangenGermany
| | - Lucas Kreiß
- Institute of Medical BiotechnologyFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 391052ErlangenGermany
- Erlangen Graduate School in Advanced Optical TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 791052ErlangenGermany
| | - Kristian Cvecek
- Erlangen Graduate School in Advanced Optical TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 791052ErlangenGermany
- Institute of Photonic TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Konrad‐Zuse‐Str. 3–591052ErlangenGermany
| | - Birgitta Carlé
- Institute of Medical BiotechnologyFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 391052ErlangenGermany
- Erlangen Graduate School in Advanced Optical TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 791052ErlangenGermany
| | - Ferdinand Knieling
- Department of Internal Medicine 1University Hospital ErlangenUlmenweg 1891054ErlangenGermany
- Department of Pediatrics and Adolescent MedicineUniversity Hospital ErlangenLoschgestr. 1591054ErlangenGermany
| | - Jean Gonzales Menezes
- Department of Internal Medicine 1University Hospital ErlangenUlmenweg 1891054ErlangenGermany
| | - Oana‐Maria Thoma
- Erlangen Graduate School in Advanced Optical TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 791052ErlangenGermany
- Department of Internal Medicine 1University Hospital ErlangenUlmenweg 1891054ErlangenGermany
| | - Michael Schmidt
- Erlangen Graduate School in Advanced Optical TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 791052ErlangenGermany
- Institute of Photonic TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Konrad‐Zuse‐Str. 3–591052ErlangenGermany
| | - Markus F. Neurath
- Erlangen Graduate School in Advanced Optical TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 791052ErlangenGermany
- Department of Internal Medicine 1University Hospital ErlangenUlmenweg 1891054ErlangenGermany
| | - Maximilian Waldner
- Erlangen Graduate School in Advanced Optical TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 791052ErlangenGermany
- Department of Internal Medicine 1University Hospital ErlangenUlmenweg 1891054ErlangenGermany
| | - Oliver Friedrich
- Institute of Medical BiotechnologyFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 391052ErlangenGermany
- Erlangen Graduate School in Advanced Optical TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 791052ErlangenGermany
| | - Sebastian Schürmann
- Institute of Medical BiotechnologyFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 391052ErlangenGermany
- Erlangen Graduate School in Advanced Optical TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 791052ErlangenGermany
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Li L, Kang D, Huang Z, Zhan Z, Feng C, Zhou Y, Tu H, Zhuo S, Chen J. Multimodal multiphoton imaging for label-free monitoring of early gastric cancer. BMC Cancer 2019; 19:295. [PMID: 30940105 PMCID: PMC6444446 DOI: 10.1186/s12885-019-5497-4] [Citation(s) in RCA: 20] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/04/2018] [Accepted: 03/20/2019] [Indexed: 12/14/2022] Open
Abstract
Background Early gastric cancer is associated with a much better prognosis than advanced disease, and strategies to improve prognosis is strictly dependent on earlier detection and accurate diagnosis. Therefore, a label-free, non-invasive imaging technique that allows the precise identification of morphologic changes in early gastric cancer would be of considerable clinical interest. Methods In this study, multiphoton microscopy (MPM) using two-photon excited fluorescence combined with second-harmonic generation was used for the identification of early gastric cancer. Results This microscope was able to directly reveal improved cellular detail and stromal changes during the development of early gastric cancer. Furthermore, two features were quantified from MPM images to assess the cell change in size and stromal collagen change as gastric lesion developed from normal to early cancer. Conclusions These results clearly show that multiphoton microscopy can be used to examine early gastric cancer at the cellular level without the need for exogenous contrast agents. This study would be helpful for early diagnosis and treatment of gastric cancer, and may provide the groundwork for further exploration into the application of multiphoton microscopy in clinical practice.
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Affiliation(s)
- Lianhuang Li
- Key Laboratory of OptoElectronic Science and Technology for Medicine of Ministry of Education, Fujian Provincial Key Laboratory for Photonics Technology, Fujian Normal University, Fuzhou, 350007, People's Republic of China.
| | - Deyong Kang
- Department of Pathology, Fujian Medical University Union Hospital, Fuzhou, 350001, People's Republic of China
| | - Zicheng Huang
- Department of Gastroenterology, The First Hospital of Quanzhou Affiliated to Fujian Medical University, Quanzhou, 362002, People's Republic of China
| | - Zhenlin Zhan
- Key Laboratory of OptoElectronic Science and Technology for Medicine of Ministry of Education, Fujian Provincial Key Laboratory for Photonics Technology, Fujian Normal University, Fuzhou, 350007, People's Republic of China
| | - Changyin Feng
- Department of Pathology, Fujian Medical University Union Hospital, Fuzhou, 350001, People's Republic of China
| | - Yongjian Zhou
- Department of Gastric Surgery, Fujian Medical University Union Hospital, Fuzhou, 350001, People's Republic of China.
| | - Haohua Tu
- Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA.
| | - Shuangmu Zhuo
- Key Laboratory of OptoElectronic Science and Technology for Medicine of Ministry of Education, Fujian Provincial Key Laboratory for Photonics Technology, Fujian Normal University, Fuzhou, 350007, People's Republic of China
| | - Jianxin Chen
- Key Laboratory of OptoElectronic Science and Technology for Medicine of Ministry of Education, Fujian Provincial Key Laboratory for Photonics Technology, Fujian Normal University, Fuzhou, 350007, People's Republic of China
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Chen D, Chen G, Jiang W, Fu M, Liu W, Sui J, Xu S, Liu Z, Zheng X, Chi L, Lin D, Li K, Chen W, Zuo N, Lu J, Chen J, Li G, Zhuo S, Yan J. Association of the Collagen Signature in the Tumor Microenvironment With Lymph Node Metastasis in Early Gastric Cancer. JAMA Surg 2019; 154:e185249. [PMID: 30698615 DOI: 10.1001/jamasurg.2018.5249] [Citation(s) in RCA: 79] [Impact Index Per Article: 13.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/06/2023]
Abstract
Importance Lymph node status is the primary determinant in treatment decision making in early gastric cancer (EGC). Current evaluation methods are not adequate for estimating lymph node metastasis (LNM) in EGC. Objective To develop and validate a prediction model based on a fully quantitative collagen signature in the tumor microenvironment to estimate the individual risk of LNM in EGC. Design, Setting, and Participants This retrospective study was conducted from August 1, 2016, to May 10, 2018, at 2 medical centers in China (Nanfang Hospital and Fujian Provincial Hospital). Participants included a primary cohort (n = 232) of consecutive patients with histologically confirmed gastric cancer who underwent radical gastrectomy and received a T1 gastric cancer diagnosis from January 1, 2008, to December 31, 2012. Patients with neoadjuvant radiotherapy, chemotherapy, or chemoradiotherapy were excluded. An additional consecutive cohort (n = 143) who received the same diagnosis from January 1, 2011, to December 31, 2013, was enrolled to provide validation. Baseline clinicopathologic data of each patient were collected. Collagen features were extracted in specimens using multiphoton imaging, and the collagen signature was constructed. An LNM prediction model based on the collagen signature was developed and was internally and externally validated. Main Outcomes and Measures The area under the receiver operating characteristic curve (AUROC) of the prediction model and decision curve were analyzed for estimating LNM. Results In total, 375 patients were included. The primary cohort comprised 232 consecutive patients, in whom the LNM rate was 16.4% (n = 38; 25 men [65.8%] with a mean [SD] age of 57.82 [10.17] years). The validation cohort consisted of 143 consecutive patients, in whom the LNM rate was 20.9% (n = 30; 20 men [66.7%] with a mean [SD] age of 54.10 [13.19] years). The collagen signature was statistically significantly associated with LNM (odds ratio, 5.470; 95% CI, 3.315-9.026; P < .001). Multivariate analysis revealed that the depth of tumor invasion, tumor differentiation, and the collagen signature were independent predictors of LNM. These 3 predictors were incorporated into the new prediction model, and a nomogram was established. The model showed good discrimination in the primary cohort (AUROC, 0.955; 95% CI, 0.919-0.991) and validation cohort (AUROC, 0.938; 95% CI, 0.897-0.981). An optimal cutoff value was selected in the primary cohort, which had a sensitivity of 86.8%, a specificity of 93.3%, an accuracy of 92.2%, a positive predictive value of 71.7%, and a negative predictive value of 97.3%. The validation cohort had a sensitivity of 90.0%, a specificity of 90.3%, an accuracy of 90.2%, a positive predictive value of 71.1%, and a negative predictive value of 97.1%. Among the 375 patients, a sensitivity of 87.3%, a specificity of 92.1%, an accuracy of 91.2%, a positive predictive value of 72.1%, and a negative predictive value of 96.9% were found. Conclusions and Relevance This study's findings suggest that the collagen signature in the tumor microenvironment is an independent indicator of LNM in EGC, and the prediction model based on this collagen signature may be useful in treatment decision making for patients with EGC.
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Affiliation(s)
- Dexin Chen
- Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, People's Republic of China.,Key Laboratory of OptoElectronic Science and Technology for Medicine of Ministry of Education, Fujian Provincial Key Laboratory of Photonics Technology, Fujian Normal University, Fuzhou, Fujian, People's Republic of China
| | - Gang Chen
- Department of Pathology, Fujian Provincial Cancer Hospital, Teaching Hospital of Fujian Medical University, Fuzhou, Fujian, People's Republic of China
| | - Wei Jiang
- Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, People's Republic of China
| | - Meiting Fu
- Department of Gastroenterology, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, People's Republic of China
| | - Wenju Liu
- Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, People's Republic of China.,Department of Pathology, Fujian Provincial Cancer Hospital, Teaching Hospital of Fujian Medical University, Fuzhou, Fujian, People's Republic of China
| | - Jian Sui
- Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, People's Republic of China.,Department of Gastrointestinal Surgery, Fujian Provincial Hospital, Teaching Hospital of Fujian Medical University, Fuzhou, Fujian, People's Republic of China
| | - Shuoyu Xu
- Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, People's Republic of China.,Department of Radiology, Sun Yat-sen University Cancer Center, Guangzhou, Guangdong, People's Republic of China
| | - Zhangyuanzhu Liu
- Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, People's Republic of China
| | - Xiaoling Zheng
- Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, People's Republic of China.,Department of Endoscopy Center, Fujian Provincial Hospital, Teaching Hospital of Fujian Medical University, Fuzhou, Fujian, People's Republic of China
| | - Liangjie Chi
- Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, People's Republic of China.,Department of Gastrointestinal Surgery, Fujian Provincial Hospital, Teaching Hospital of Fujian Medical University, Fuzhou, Fujian, People's Republic of China
| | - Dajia Lin
- Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, People's Republic of China.,Department of Gastrointestinal Surgery, Fujian Provincial Hospital, Teaching Hospital of Fujian Medical University, Fuzhou, Fujian, People's Republic of China
| | - Kai Li
- Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, People's Republic of China
| | - Weisheng Chen
- Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, People's Republic of China
| | - Ning Zuo
- Key Laboratory of OptoElectronic Science and Technology for Medicine of Ministry of Education, Fujian Provincial Key Laboratory of Photonics Technology, Fujian Normal University, Fuzhou, Fujian, People's Republic of China
| | - Jianping Lu
- Department of Pathology, Fujian Provincial Cancer Hospital, Teaching Hospital of Fujian Medical University, Fuzhou, Fujian, People's Republic of China
| | - Jianxin Chen
- Key Laboratory of OptoElectronic Science and Technology for Medicine of Ministry of Education, Fujian Provincial Key Laboratory of Photonics Technology, Fujian Normal University, Fuzhou, Fujian, People's Republic of China
| | - Guoxin Li
- Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, People's Republic of China
| | - Shuangmu Zhuo
- Key Laboratory of OptoElectronic Science and Technology for Medicine of Ministry of Education, Fujian Provincial Key Laboratory of Photonics Technology, Fujian Normal University, Fuzhou, Fujian, People's Republic of China
| | - Jun Yan
- Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, People's Republic of China
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MEMS Actuators for Optical Microendoscopy. MICROMACHINES 2019; 10:mi10020085. [PMID: 30682852 PMCID: PMC6412441 DOI: 10.3390/mi10020085] [Citation(s) in RCA: 14] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 11/28/2018] [Revised: 12/26/2018] [Accepted: 12/28/2018] [Indexed: 01/21/2023]
Abstract
Growing demands for affordable, portable, and reliable optical microendoscopic imaging devices are attracting research institutes and industries to find new manufacturing methods. However, the integration of microscopic components into these subsystems is one of today's challenges in manufacturing and packaging. Together with this kind of miniaturization more and more functional parts have to be accommodated in ever smaller spaces. Therefore, solving this challenge with the use of microelectromechanical systems (MEMS) fabrication technology has opened the promising opportunities in enabling a wide variety of novel optical microendoscopy to be miniaturized. MEMS fabrication technology enables abilities to apply batch fabrication methods with high-precision and to include a wide variety of optical functionalities to the optical components. As a result, MEMS technology has enabled greater accessibility to advance optical microendoscopy technology to provide high-resolution and high-performance imaging matching with traditional table-top microscopy. In this review the latest advancements of MEMS actuators for optical microendoscopy will be discussed in detail.
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Seo YH, Hwang K, Kim H, Jeong KH. Scanning MEMS Mirror for High Definition and High Frame Rate Lissajous Patterns. MICROMACHINES 2019; 10:mi10010067. [PMID: 30669314 PMCID: PMC6356757 DOI: 10.3390/mi10010067] [Citation(s) in RCA: 15] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 12/04/2018] [Revised: 01/16/2019] [Accepted: 01/17/2019] [Indexed: 11/16/2022]
Abstract
Scanning MEMS (micro-electro-mechanical system) mirrors are attractive given their potential use in a diverse array of laser scanning display and imaging applications. Here we report on an electrostatic MEMS mirror for high definition and high frame rate (HDHF) Lissajous scanning. The MEMS mirror comprised a low Q-factor inner mirror and frame mirror, which provided two-dimensional scanning at two similar resonant scanning frequencies with high mechanical stability. The low Q inner mirror enabled a broad frequency selection range. The high definition and high frame rate (HDHF) Lissajous scanning of the MEMS mirror was achieved by selecting a set of scanning frequencies near its resonance with a high greatest common divisor (GCD) and a high total lobe number. The MEMS mirror had resonant scanning frequencies at 5402 Hz and 6702 Hz in x and y directions, respectively. The selected pseudo-resonant frequencies of 5450 Hz and 6700 Hz for HDHF scanning provided 50 frames per second with 94% fill factor in 256 × 256 pixels. This Lissajous MEMS mirror could be utilized for assorted HDHF laser scanning imaging and display applications.
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Affiliation(s)
- Yeong-Hyeon Seo
- Department of Bio and Brain Engineering, KAIST, Daejeon 34141, Korea.
- KAIST Institute for Health Science and Technology, Daejeon 34141, Korea.
| | - Kyungmin Hwang
- Department of Bio and Brain Engineering, KAIST, Daejeon 34141, Korea.
- KAIST Institute for Health Science and Technology, Daejeon 34141, Korea.
| | - Hyunwoo Kim
- Department of Bio and Brain Engineering, KAIST, Daejeon 34141, Korea.
- KAIST Institute for Health Science and Technology, Daejeon 34141, Korea.
| | - Ki-Hun Jeong
- Department of Bio and Brain Engineering, KAIST, Daejeon 34141, Korea.
- KAIST Institute for Health Science and Technology, Daejeon 34141, Korea.
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Saitou T, Takanezawa S, Ninomiya H, Watanabe T, Yamamoto S, Hiasa Y, Imamura T. Tissue Intrinsic Fluorescence Spectra-Based Digital Pathology of Liver Fibrosis by Marker-Controlled Segmentation. Front Med (Lausanne) 2019; 5:350. [PMID: 30619861 PMCID: PMC6297145 DOI: 10.3389/fmed.2018.00350] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/18/2018] [Accepted: 11/28/2018] [Indexed: 01/16/2023] Open
Abstract
Tissue intrinsic emission fluorescence provides useful diagnostic information for various diseases. Because of its unique feature of spectral profiles depending on tissue types, spectroscopic imaging is a promising tool for accurate evaluation of endogenous fluorophores. However, due to difficulties in discriminating those sources, quantitative analysis remains challenging. In this study, we quantitatively investigated spectral-spatial features of multi-photon excitation fluorescence in normal and diseased livers. For morphometrics of multi-photon excitation spectra, we examined a marker-controlled segmentation approach and its application to liver fibrosis assessment by employing a mouse model of carbon tetrachloride (CCl4)-induced liver fibrosis. We formulated a procedure of internal marker selection where markers were chosen to reflect typical biochemical species in the liver, followed by image segmentation and local morphological feature extraction. Image segmentation enabled us to apply mathematical morphology analysis, and the local feature was applied to the automated classification test based on a machine learning framework, both demonstrating highly accurate classifications. Through the analyses, we showed that spectral imaging of native fluorescence from liver tissues have the capability of differentiating not only between normal and diseased, but also between progressive disease states. The proposed approach provides the basics of spectroscopy-based digital histopathology of chronic liver diseases, and can be applied to a range of diseases associated with autofluorescence alterations.
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Affiliation(s)
- Takashi Saitou
- Department of Molecular Medicine for Pathogenesis, Graduate School of Medicine, Ehime University, Toon, Japan.,Translational Research Center, Ehime University Hospital, Toon, Japan.,Division of Bio-Imaging, Proteo-Science Center (PROS), Ehime University, Toon, Japan
| | - Sota Takanezawa
- Department of Molecular Medicine for Pathogenesis, Graduate School of Medicine, Ehime University, Toon, Japan
| | - Hiroko Ninomiya
- Department of Molecular Medicine for Pathogenesis, Graduate School of Medicine, Ehime University, Toon, Japan
| | - Takao Watanabe
- Department of Gastroenterology and Metabiology, Graduate School of Medicine, Ehime University, Toon, Japan
| | - Shin Yamamoto
- Department of Gastroenterology and Metabiology, Graduate School of Medicine, Ehime University, Toon, Japan.,Department of Lifestyle-related Medicine and Endocrinology, Graduate School of Medicine, Ehime University, Toon, Japan
| | - Yoichi Hiasa
- Department of Gastroenterology and Metabiology, Graduate School of Medicine, Ehime University, Toon, Japan
| | - Takeshi Imamura
- Department of Molecular Medicine for Pathogenesis, Graduate School of Medicine, Ehime University, Toon, Japan.,Translational Research Center, Ehime University Hospital, Toon, Japan.,Division of Bio-Imaging, Proteo-Science Center (PROS), Ehime University, Toon, Japan
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47
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Prasad A, Chaichi A, Kelley DP, Francis J, Gartia MR. Current and future functional imaging techniques for post-traumatic stress disorder. RSC Adv 2019; 9:24568-24594. [PMID: 35527877 PMCID: PMC9069787 DOI: 10.1039/c9ra03562a] [Citation(s) in RCA: 13] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/12/2019] [Accepted: 08/02/2019] [Indexed: 11/21/2022] Open
Abstract
Posttraumatic stress disorder (PTSD) is a trauma and stressor related psychiatric disorder associated with structural, metabolic, and molecular alternations in several brain regions including diverse cortical areas, neuroendocrine regions, the striatum, dopaminergic, adrenergic and serotonergic pathways, and the limbic system. We are in critical need of novel therapeutics and biomarkers for PTSD and a deep understanding of cutting edge imaging and spectroscopy methods is necessary for the development of promising new approaches to better diagnose and treat the disorder. According to the Diagnostic and Statistical Manual of Mental Disorders (DSM-V) criterion, all forms of traumatic stress-induced disorder are considered acute stress disorder for the first month following the stressor. Only after symptoms do not remit for one month can the disorder be deemed PTSD. It would be particularly useful to differentiate between acute stress disorder and PTSD during the one month waiting period so that more intensive treatments can be applied early on to patients with a high likelihood of developing PTSD. This would potentially enhance treatment outcomes and/or prevent the development of PTSD. Comprehension of the qualities and limitations of currently applied methods as well as the novel emerging techniques provide invaluable knowledge for fast paced development. Conventional methods of studying PTSD have proven to be insufficient for diagnosis, measurement of treatment efficacy, and monitoring disease progression. As the field currently stands, there is no diagnostic biomarker available for any psychiatric disease, PTSD included. Currently, emerging and available technologies are not utilized to their full capacity and in appropriate experimental designs for the most fruitful possible studies in this area. Therefore, there is an apparent need for improved methods in PTSD research. This review demonstrates the current state of the literature in PTSD, including molecular, cellular, and behavioral indicators, possible biomarkers and clinical and pre-clinical imaging techniques relevant to PTSD, and through this, elucidate the void of current practical imaging and spectroscopy methods that provide true biomarkers for the disorder and the significance of devising new techniques for future investigations. We are unlikely to develop a single biomarker for any psychiatric disorder however. As psychiatric disorders are incomparably complex compared to other medical diagnoses, its most likely that transcriptomic, metabolomic and structural and connectomic imaging data will have to be analyzed in concert in order to produce a dependable non-behavioral marker of PTSD. This can explain the necessity of bridging conventional approaches to novel technologies in order to create a framework for further discoveries in the treatment of PTSD. Conventional methods of studying posttraumatic stress disorder (PTSD) have proven to be insufficient for diagnosis. We have reviewed clinical and preclinical imaging techniques as well as molecular, cellular, and behavioral indicators for PTSD.![]()
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Affiliation(s)
- Alisha Prasad
- Department of Mechanical and Industrial Engineering
- Louisiana State University
- Baton Rouge
- USA
| | - Ardalan Chaichi
- Department of Mechanical and Industrial Engineering
- Louisiana State University
- Baton Rouge
- USA
| | - D. Parker Kelley
- Comparative Biomedical Sciences
- School of Veterinary Medicine
- Louisiana State University
- Baton Rouge
- USA
| | - Joseph Francis
- Comparative Biomedical Sciences
- School of Veterinary Medicine
- Louisiana State University
- Baton Rouge
- USA
| | - Manas Ranjan Gartia
- Department of Mechanical and Industrial Engineering
- Louisiana State University
- Baton Rouge
- USA
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48
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Baskey SJ, Andreana M, Lanteigne E, Ridsdale A, Stolow A, Schweitzer ME. Pre-Clinical Translation of Second Harmonic Microscopy of Meniscal and Articular Cartilage Using a Prototype Nonlinear Microendoscope. IEEE JOURNAL OF TRANSLATIONAL ENGINEERING IN HEALTH AND MEDICINE-JTEHM 2018; 7:1800211. [PMID: 30701146 PMCID: PMC6342420 DOI: 10.1109/jtehm.2018.2889496] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 08/16/2018] [Revised: 11/05/2018] [Accepted: 11/30/2018] [Indexed: 11/06/2022]
Abstract
Previous studies using nonlinear microscopy have demonstrated that osteoarthritis (OA) is characterized by the gradual replacement of Type II collagen with Type I collagen. The objective of this study was to develop a prototype nonlinear laser scanning microendoscope capable of resolving the structural differences of collagen in various orthopaedically relevant cartilaginous surfaces. The current prototype developed a miniaturized femtosecond laser scanning instrument, mounted on an articulated positioning system, capable of both conventional arthroscopy and second-harmonic laser-scanning microscopy. Its optical system includes a multi-resolution optical system using a gradient index objective lens and a customized multi-purpose fiber optic sheath to maximize the collection of backscattered photons or provide joint capsule illumination. The stability and suitability of the prototype arthroscope to approach and image cartilage were evaluated through preliminary testing on fresh, minimally processed, and partially intact porcine knee joints. Image quality was sufficient to distinguish between hyaline cartilage and fibrocartilage through unique Type I and Type II collagen-specific characteristics. Imaging the meniscus revealed that the system was able to visualize differences in the collagen arrangement between the superficial and lamellar layers. Such detailed in vivo imaging of the cartilage surfaces could obviate the need to perform biopsies for ex vivo histological analysis in the future, and provide an alternative to conventional external imaging to characterize and diagnose progressive and degenerative cartilage diseases such as OA. Moreover, this system is readily customizable and may provide a suitable and modular platform for developing additional tools utilizing femtosecond lasers for tissue cutting within the familiar confines of two or three portal arthroscopy techniques.
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Affiliation(s)
- Stephen J Baskey
- Faculty of MedicineUniversity of OttawaOttawaONK1H 8M5Canada.,Department of Mechanical EngineeringUniversity of OttawaOttawaONK1N 6N5Canada.,Emerging Technologies Division, Molecular Photonics GroupNational Research Council CanadaOttawaONK1A 0R6Canada
| | - Marco Andreana
- Center for Medical Physics and Biomedical EngineeringMedical University of Vienna1090ViennaAustria
| | - Eric Lanteigne
- Department of Mechanical EngineeringUniversity of OttawaOttawaONK1N 6N5Canada
| | - Andrew Ridsdale
- Emerging Technologies Division, Molecular Photonics GroupNational Research Council CanadaOttawaONK1A 0R6Canada
| | - Albert Stolow
- Emerging Technologies Division, Molecular Photonics GroupNational Research Council CanadaOttawaONK1A 0R6Canada.,Department of PhysicsUniversity of OttawaOttawaONK1N 6N5Canada.,Department of ChemistryUniversity of OttawaOttawaONK1N 6N5Canada
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49
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Huang L, Zhou X, Tang S. Optimization of frequency-doubled Er-doped fiber laser for miniature multiphoton endoscopy. JOURNAL OF BIOMEDICAL OPTICS 2018; 23:1-12. [PMID: 30574695 DOI: 10.1117/1.jbo.23.12.126503] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 06/05/2018] [Accepted: 11/26/2018] [Indexed: 05/18/2023]
Abstract
Frequency-doubled femtosecond Er-doped fiber laser is a low-cost and portable excitation source suitable for multiphoton endoscopy. The frequency-doubled wavelength at 780 nm is used to excite the intrinsic fluorescence signal. The frequency-doubling with a periodically poled MgO : LiNbO3 (PPLN) is integrated in the distal end of the imaging head to achieve fiber connection. The imaging speed is further improved by optimizing the excitation laser source. A 0.3-mm length of PPLN crystal is selected and the Er-doped fiber laser is manipulated to match its bandwidth with the acceptance bandwidth of the PPLN. Through this optimization, a reduced pulsewidth of 80 fs of the frequency-doubled pulse is achieved. All-fiber dispersion compensation and pulse compression by single mode fiber is conducted, which makes the fiber laser directly fiber-coupled to the imaging head. An imaging speed of 4 frames / s is demonstrated on ex vivo imaging of unstained biological tissues, which is 10 times faster than our previous study using a 1-mm-long PPLN. The results show that miniature multiphoton endoscopy using frequency-doubled Er-doped fiber laser has great potential for clinical applications.
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Affiliation(s)
- Lin Huang
- University of British Columbia, Department of Electrical and Computer Engineering, Vancouver, Canada
| | - Xin Zhou
- University of British Columbia, Department of Electrical and Computer Engineering, Vancouver, Canada
| | - Shuo Tang
- University of British Columbia, Department of Electrical and Computer Engineering, Vancouver, Canada
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50
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Liang W, Hall G, Li X. Spectro-temporal dispersion management of femtosecond pulses for fiber-optic two-photon endomicroscopy. OPTICS EXPRESS 2018; 26:22877-22893. [PMID: 30184945 PMCID: PMC6238828 DOI: 10.1364/oe.26.022877] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 06/08/2018] [Revised: 07/26/2018] [Accepted: 07/27/2018] [Indexed: 05/19/2023]
Abstract
The emerging fiber-optic two-photon endomicroscopy technology holds a strong promise for enabling translational applications of nonlinear optical imaging. Effective femtosecond pulse dispersion management is critical for achieving high-quality imaging. Here we report systematic analyses and performance characterization of a dual-fiber spectro-temporal dispersion management scheme involving a grating pair as the pulse stretcher. Compared with conventional linear-only compensation, the grating-based spectro-temporal compensation also takes into account nonlinear effects and enhances the two-photon signal by ~3-fold as experimentally demonstrated. Numerical simulations were carried out to systematically investigate the influence of several key design parameters on the overall compensation efficacy. Furthermore, comprehensive performance comparison with an ideal grism-pair counterpart reveals that a grating-pair stretcher affords much higher power throughput and thus is preferable for portable endomicroscopy systems with limited laser source power.
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Affiliation(s)
- Wenxuan Liang
- Department of Biomedical Engineering, Johns Hopkins University, 720 Rutland Ave, Baltimore, Maryland, 21205, USA
| | - Gunnsteinn Hall
- Department of Biomedical Engineering, Johns Hopkins University, 720 Rutland Ave, Baltimore, Maryland, 21205, USA
| | - Xingde Li
- Department of Biomedical Engineering, Johns Hopkins University, 720 Rutland Ave, Baltimore, Maryland, 21205, USA
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