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Clarke KSP, Lewis R, Hughes MP, Labeed FH. Tracking chondrocyte-to-fibroblast transformation via changes in cell electrophysiology. Sci Rep 2025; 15:15865. [PMID: 40335587 PMCID: PMC12059058 DOI: 10.1038/s41598-025-98958-1] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/10/2024] [Accepted: 04/16/2025] [Indexed: 05/09/2025] Open
Abstract
Cultured chondrocytes have potential for cartilage regeneration to treat degenerative diseases. However, when explanted chondrocytes are cultured in monolayer, they are known to dedifferentiate over time, adopting a more fibroblastic phenotype. This greatly impacts both research and potential clinical applications; cell-based cartilage repair therapies require significant in vitro expansion to obtain sufficient chondrocyte numbers for reimplantation, as chondrocytes adopt a more fibroblastic phenotype causing up to 70% of patients to develop fibrocartilaginous fill. We used dielectrophoresis (DEP) to observe changes in the electrophysiological properties of primary bovine chondrocytes over time. Using a multi-conductivity approach, we demonstrate that monitoring the cytoplasmic conductivity is a reliable method of observing cell changes over 100 days in culture. Results show statistically significant changes in both membrane capacitance (p = 0.0039) and cytoplasm conductivity (p = < 0.0001) when tested at multiple conductivities. Analysis of cytoplasmic vs. medium conductivity allowed simple tracking of chondrocyte membrane potential, which exhibited transitions between three stable values of Vm which correspond to patch-clamp-derived literature values as they transition from chondrocytes (- 13 to - 18 mV) to proliferating fibroblasts (- 32 to - 43 mV) and ultimately to non-proliferating fibroblasts (- 55 to - 71 mV), transitions occurring around days 40 and 80 respectively.
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Affiliation(s)
- Krista S P Clarke
- Centre for Biomedical Engineering, School of Mechanical Engineering, University of Surrey, Guildford, Surrey, GU2 7XH, UK
| | - Rebecca Lewis
- Department of Comparative Biomedical Sciences, School of Veterinary Medicine, University of Surrey, Guildford, Surrey, GU2 7XH, UK
| | - Michael Pycraft Hughes
- Centre for Biomedical Engineering, School of Mechanical Engineering, University of Surrey, Guildford, Surrey, GU2 7XH, UK
- Department of Biomedical Engineering and Biotechnology, Khalifa University, Abu Dhabi, United Arab Emirates
| | - Fatima H Labeed
- Centre for Biomedical Engineering, School of Mechanical Engineering, University of Surrey, Guildford, Surrey, GU2 7XH, UK.
- Department of Biology, United Arab Emirates University, Al Ain, United Arab Emirates.
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Vaghef-Koodehi A, Lapizco-Encinas BH. Dielectrophoresis in Carcinoma Diagnosis: Recent Developments and Applications. Electrophoresis 2025. [PMID: 40099708 DOI: 10.1002/elps.8127] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/18/2024] [Revised: 01/07/2025] [Accepted: 02/20/2025] [Indexed: 03/20/2025]
Abstract
Accurate cancer cell analysis is critical across a wide range of medical fields, including clinical diagnostics, personalized medicine, drug development, and cancer research. The ability to rapidly analyze and characterize cancer cells is key to understanding tumor characteristics, developing targeted therapies, and improving patient outcomes. Microscale electrokinetic (EK) techniques have demonstrated their effectiveness and reliability as powerful tools for cell analysis, including cancerous cells. The applications of dielectrophoresis (DEP), an EK technique, in cancer cell analysis are discussed here with a focus on carcinomas, cancer that develops in epithelial tissue. In this review article, the working mechanism of DEP is first introduced, followed by an in-depth review of recent studies published between 2015 and 2024. The concluding remarks section provides a summary of the key points discussed in this review and offers insights into potential future advancements in DEP-based systems for analyzing cancerous cells.
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Affiliation(s)
- Alaleh Vaghef-Koodehi
- Microscale Bioseparations Laboratory and Biomedical Engineering Department, Rochester Institute of Technology, Rochester, New York, USA
| | - Blanca H Lapizco-Encinas
- Microscale Bioseparations Laboratory and Biomedical Engineering Department, Rochester Institute of Technology, Rochester, New York, USA
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Mathews J, Erickson P, Kuchling F, Jawanda N, Pio-Lopez L, Pai VP, Levin M. Meeting Review: "National Cancer Institute Conference on Cancer Bioelectricity" September 12, 2024. Bioelectricity 2025; 7:94-104. [PMID: 40342939 PMCID: PMC12054611 DOI: 10.1089/bioe.2024.0049] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 05/11/2025] Open
Abstract
The Office of Cancer Complementary and Alternative Medicine, part of the Division of Cancer Treatment and Diagnosis under the National Cancer Institute (NCI), supports research in diverse areas of cancer therapeutics such as microbial therapies, herbal remedies, and mind-body practices. Recently they have become especially interested in the emerging role of bioelectricity in cancer biology and organized a virtual meeting with some of the top scientists in the field. In this report, we overview this first-of-its-kind Naional Institute of Health (NIH)-sponsored meeting, which featured talks from 14 researchers exploring the role of bioelectricity in cancer biology. The talks covered a wide range of topics, including excellent background information on how cell collectives change their bioelectrical coupling and set points during cancer formation, new tools for reading and writing bioelectrical signatures in cells and whole organisms, how ion channels that are involved in setting those signatures affect canonical pathways in development and tumor growth, and the methods for modeling bioelectrical interactions and information transfer in cell collectives. Especially exciting were the translational technologies that were highlighted, including new diagnostics, metastasis inhibition therapies, and more efficient detection of surgical margins. The meeting concluded with funding opportunities available from the NCI Division of Cancer Biology, Innovative Molecular Analysis Technologies Program, and the Small Business Innovation Research Development Center.
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Affiliation(s)
- Juanita Mathews
- Allen Discovery Center at Tufts University, Medford, Massachusetts, USA
| | - Patrick Erickson
- Allen Discovery Center at Tufts University, Medford, Massachusetts, USA
| | - Franz Kuchling
- Allen Discovery Center at Tufts University, Medford, Massachusetts, USA
| | - Navneet Jawanda
- Allen Discovery Center at Tufts University, Medford, Massachusetts, USA
| | - Léo Pio-Lopez
- Allen Discovery Center at Tufts University, Medford, Massachusetts, USA
| | - Vaibhav P. Pai
- Allen Discovery Center at Tufts University, Medford, Massachusetts, USA
| | - Michael Levin
- Allen Discovery Center at Tufts University, Medford, Massachusetts, USA
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Hoque R, Labeed FH, Cirovic S, Hughes MP. Evaluating refrigeration and antibiotic treatment for maintaining urine electrophysiology. PLoS One 2025; 20:e0319089. [PMID: 39999201 PMCID: PMC11856264 DOI: 10.1371/journal.pone.0319089] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/19/2024] [Accepted: 01/28/2025] [Indexed: 02/27/2025] Open
Abstract
BACKGROUND Electrophysiological analysis of urine has shown utility in differentiating between healthy and bladder cancer specimens, offering a rapid, label-free alternative to molecular methods. However, transporting and preserving urine samples from collection to the laboratory poses logistical challenges that could impact the reliability of electrophysiological measurements. OBJECTIVE This study investigates the effects of prolonged refrigeration on the dielectric properties and ζ-potential of urine specimens and evaluate whether antibiotic treatment can enhance sample preservation without altering electrophysiological properties. A new methodology to evaluate urine specimen quality and determine bacterial contamination, using electrophysiological modalities, is presented. METHODS Mid-stream urine samples from healthy participants (n = 4) were collected and divided into untreated and 1% penicillin/streptomycin-treated groups. Samples were analysed at baseline prior to storage at 4°C, with further analysis every 24 hours for 96 hours. Changes in dielectrophoresis (DEP) response and ζ-potential were measured using a 3DEP cytometer (Deparator, UK) and Malvern Panalytical Zetasizer Nano ZS90 (Malvern, UK), respectively. Chemical analyses, including pH and nitrite levels, and microscopic examinations were also conducted. RESULTS & LIMITATIONS Significant electrophysiological changes were observed in both untreated and antibiotic-treated urine samples over time. Both groups showed a linear increase of change in DEP response and ζ-potential values, from baseline over time. Untreated samples exhibited significant deviations in DEP and ζ-potential from baseline after 48 hours, with significance at 72 hours (P < 0.05). Treated samples only showed significant changes in ζ-potential at 96 hours (P < 0.05). Chemical analysis indicated increased pH and nitrite presence in untreated samples at 48 hours, indicating bacterial growth. Treated samples took more than 48 hours to show changes in both chemical parameters. Limitations include the relatively small study sample size, not evaluating the preservatory effects of UTI-specific antibiotics, such as nitrofurantoin and trimethoprim, and exploring different drug concentrations. CONCLUSION Prolonged refrigeration can maintain the quality of urine samples for up to 48 hours with antibiotic treatment. Current UK and European guidelines recommend urinalysis within 24 hours of specimen collection; the findings of this study support the use of DEP and ζ-potential analysis as practical clinical tests in a mail-in screening setting, provided appropriate sample preservation measures are taken.
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Affiliation(s)
- Rashedul Hoque
- Centre for Biomedical Engineering, School of Engineering, University of Surrey, Guildford, United Kingdom
| | - Fatima H. Labeed
- Department of Biology, UAEU University, Al Ain, United Arab Emirates
| | - Srdjan Cirovic
- Centre for Biomedical Engineering, School of Engineering, University of Surrey, Guildford, United Kingdom
| | - Michael Pycraft Hughes
- Department of Biomedical Engineering and Biotechnology/Healthcare Engineering Innovation Group, Khalifa University of Science and Technology, Abu Dhabi, United Arab Emirates
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Clarke KSP, Griffiths OV, Hoettges K, Lewis R, Labeed FH. Quantifying the Electrophysiological Properties of Cells Using Dielectrophoresis. Methods Mol Biol 2025; 2908:191-209. [PMID: 40304911 DOI: 10.1007/978-1-0716-4434-8_13] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 05/02/2025]
Abstract
The 3DEP Reader (DEPtech) is a commercial noninvasive, high throughput, easily adoptable benchtop microfluidic platform that utilizes the principles of dielectrophoresis to analyze cells in suspension and determine their electrical properties. This protocol describes the operation, value, and common setup of the platform to provide users with the ability to characterize three important electrophysiological parameters of approximately 20,000 cells in under 2 min. Dielectrophoresis (DEP) is an electrokinetic phenomenon which describes the movement of suspended polarizable cells, in a nonuniform electric field. The 3DEP plots the relative DEP force experienced by cells across a range of AC frequencies generating a spectrum through which membrane conductance, membrane capacitance, and cytoplasmic conductivity are derived.
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Affiliation(s)
- Krista S P Clarke
- Centre for Biomedical Engineering, School of Mechanical Engineering Sciences, University of Surrey, Guildford, Surrey, UK
| | - Oreoluwa V Griffiths
- Centre for Biomedical Engineering, School of Mechanical Engineering Sciences, University of Surrey, Guildford, Surrey, UK
| | - Kai Hoettges
- Department of Electrical Engineering and Electronics, University of Liverpool, Liverpool, UK
| | - Rebecca Lewis
- Department of Comparative Biomedical Sciences, School of Veterinary Medicine, University of Surrey, Guildford, Surrey, UK
| | - Fatima H Labeed
- Department of Biology, United Arab Emirates University, Al Ain, UAE.
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Hughes MP, Clarke KSP, Hoque R, Griffiths OV, Kruchek EJ, Johnson MP, Tariq MH, Kohli N, Lewis R, Labeed FH. Label-free, non-contact determination of resting membrane potential using dielectrophoresis. Sci Rep 2024; 14:18477. [PMID: 39122771 PMCID: PMC11316104 DOI: 10.1038/s41598-024-69000-7] [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] [Received: 03/06/2024] [Accepted: 07/30/2024] [Indexed: 08/12/2024] Open
Abstract
Measurement of cellular resting membrane potential (RMP) is important in understanding ion channels and their role in regulation of cell function across a wide range of cell types. However, methods available for the measurement of RMP (including patch clamp, microelectrodes, and potential-sensitive fluorophores) are expensive, slow, open to operator bias, and often result in cell destruction. We present non-contact, label-free membrane potential estimation which uses dielectrophoresis to determine the cytoplasm conductivity slope as a function of medium conductivity. By comparing this to patch clamp data available in the literature, we have demonstratet the accuracy of this approach using seven different cell types, including primary suspension cells (red blood cells, platelets), cultured suspension cells (THP-1), primary adherent cells (chondrocytes, human umbilical mesenchymal stem cells), and adherent (HeLa) and suspension (Jurkat) cancer cell lines. Analysis of the effect of ion channel inhibitors suggests the effects of pharmaceutical agents (TEA on HeLa; DMSO and neuraminidase on red blood cells) can also be measured. Comparison with published values of membrane potential suggest that the differences between our estimates and values recorded by patch clamp are accurate to within published margins of error. The method is low-cost, non-destructive, operator-independent and label-free, and has previously been shown to allow cells to be recovered after measurement.
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Affiliation(s)
- Michael Pycraft Hughes
- Department of Biomedical Engineering and Biotechnology, Khalifa University of Science and Technology, Abu Dhabi, UAE.
- Healthcare Engineering Innovation Center, Khalifa University of Science and Technology, Abu Dhabi, UAE.
| | - Krista S P Clarke
- Centre for Biomedical Engineering, University of Surrey, Guildford, Surrey, GU2 7XH, UK
| | - Rashedul Hoque
- Centre for Biomedical Engineering, University of Surrey, Guildford, Surrey, GU2 7XH, UK
| | - Oreoluwa V Griffiths
- Centre for Biomedical Engineering, University of Surrey, Guildford, Surrey, GU2 7XH, UK
| | - Emily J Kruchek
- Centre for Biomedical Engineering, University of Surrey, Guildford, Surrey, GU2 7XH, UK
| | - Matthew P Johnson
- Department of Biomedical Engineering and Biotechnology, Khalifa University of Science and Technology, Abu Dhabi, UAE
| | - Muhammad Hamza Tariq
- Department of Biomedical Engineering and Biotechnology, Khalifa University of Science and Technology, Abu Dhabi, UAE
| | - Nupur Kohli
- Department of Biomedical Engineering and Biotechnology, Khalifa University of Science and Technology, Abu Dhabi, UAE
- Healthcare Engineering Innovation Center, Khalifa University of Science and Technology, Abu Dhabi, UAE
| | - Rebecca Lewis
- Department of Comparative Biomedical Sciences, School of Veterinary Medicine, University of Surrey, Guildford, Surrey, GU2 7XH, UK
| | - Fatima H Labeed
- Department of Biology, United Arab Emirates University, Al Ain, UAE.
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7
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Dahal N, Peak C, Ehrett C, Osterberg J, Cao M, Divan R, Wang P. Microwave Flow Cytometric Detection and Differentiation of Escherichia coli. SENSORS (BASEL, SWITZERLAND) 2024; 24:2870. [PMID: 38732977 PMCID: PMC11086155 DOI: 10.3390/s24092870] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 02/19/2024] [Revised: 04/24/2024] [Accepted: 04/26/2024] [Indexed: 05/13/2024]
Abstract
Label-free measurement and analysis of single bacterial cells are essential for food safety monitoring and microbial disease diagnosis. We report a microwave flow cytometric sensor with a microstrip sensing device with reduced channel height for bacterial cell measurement. Escherichia coli B and Escherichia coli K-12 were measured with the sensor at frequencies between 500 MHz and 8 GHz. The results show microwave properties of E. coli cells are frequency-dependent. A LightGBM model was developed to classify cell types at a high accuracy of 0.96 at 1 GHz. Thus, the sensor provides a promising label-free method to rapidly detect and differentiate bacterial cells. Nevertheless, the method needs to be further developed by comprehensively measuring different types of cells and demonstrating accurate cell classification with improved machine-learning techniques.
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Affiliation(s)
- Neelima Dahal
- Holcombe Department of Electrical and Computer Engineering, Clemson University, Clemson, SC 29634, USA; (N.D.); (J.O.)
| | - Caroline Peak
- Department of Bioengineering, Clemson University, Clemson, SC 29634, USA;
| | - Carl Ehrett
- Watt Family Innovation Center, Clemson University, Clemson, SC 29634, USA;
| | - Jeffrey Osterberg
- Holcombe Department of Electrical and Computer Engineering, Clemson University, Clemson, SC 29634, USA; (N.D.); (J.O.)
| | - Min Cao
- Department of Biological Sciences, Clemson University, Clemson, SC 29634, USA;
| | - Ralu Divan
- Argonne National Laboratory, Chicago, IL 60439, USA;
| | - Pingshan Wang
- Holcombe Department of Electrical and Computer Engineering, Clemson University, Clemson, SC 29634, USA; (N.D.); (J.O.)
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Borenstein JT, Cummins G, Dutta A, Hamad E, Hughes MP, Jiang X, Lee HH, Lei KF, Tang XS, Zheng Y, Chen J. Bionanotechnology and bioMEMS (BNM): state-of-the-art applications, opportunities, and challenges. LAB ON A CHIP 2023; 23:4928-4949. [PMID: 37916434 DOI: 10.1039/d3lc00296a] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 11/03/2023]
Abstract
The development of micro- and nanotechnology for biomedical applications has defined the cutting edge of medical technology for over three decades, as advancements in fabrication technology developed originally in the semiconductor industry have been applied to solving ever-more complex problems in medicine and biology. These technologies are ideally suited to interfacing with life sciences, since they are on the scale lengths as cells (microns) and biomacromolecules (nanometers). In this paper, we review the state of the art in bionanotechnology and bioMEMS (collectively BNM), including developments and challenges in the areas of BNM, such as microfluidic organ-on-chip devices, oral drug delivery, emerging technologies for managing infectious diseases, 3D printed microfluidic devices, AC electrokinetics, flexible MEMS devices, implantable microdevices, paper-based microfluidic platforms for cellular analysis, and wearable sensors for point-of-care testing.
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Affiliation(s)
| | - Gerard Cummins
- School of Engineering, University of Birmingham, Edgbaston, B15 2TT, UK.
| | - Abhishek Dutta
- Department of Electrical & Computer Engineering, University of Connecticut, USA.
| | - Eyad Hamad
- Biomedical Engineering Department, School of Applied Medical Sciences, German Jordanian University, Amman, Jordan.
| | - Michael Pycraft Hughes
- Department of Biomedical Engineering, Khalifa University, Abu Dhabi, United Arab Emirates.
| | - Xingyu Jiang
- Department of Biomedical Engineering, Southern University of Science and Technology, China.
| | - Hyowon Hugh Lee
- Weldon School of Biomedical Engineering, Center for Implantable Devices, Purdue University, West Lafayette, IN, USA.
| | | | | | | | - Jie Chen
- Department of Electrical and Computer Engineering, University of Alberta, Edmonton, AB T6G 2R3, Canada.
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Deivasigamani R, Mohd Maidin NN, Abdul Nasir NS, Abdulhameed A, Ahmad Kayani AB, Mohamed MA, Buyong MR. A correlation of conductivity medium and bioparticle viability on dielectrophoresis-based biomedical applications. Electrophoresis 2023; 44:573-620. [PMID: 36604943 DOI: 10.1002/elps.202200203] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/12/2022] [Revised: 11/28/2022] [Accepted: 12/26/2022] [Indexed: 01/07/2023]
Abstract
Dielectrophoresis (DEP) bioparticle research has progressed from micro to nano levels. It has proven to be a promising and powerful cell manipulation method with an accurate, quick, inexpensive, and label-free technique for therapeutic purposes. DEP, an electrokinetic phenomenon, induces particle movement as a result of polarization effects in a nonuniform electrical field. This review focuses on current research in the biomedical field that demonstrates a practical approach to DEP in terms of cell separation, trapping, discrimination, and enrichment under the influence of the conductive medium in correlation with bioparticle viability. The current review aims to provide readers with an in-depth knowledge of the fundamental theory and principles of the DEP technique, which is influenced by conductive medium and to identify and demonstrate the biomedical application areas. The high conductivity of physiological fluids presents obstacles and opportunities, followed by bioparticle viability in an electric field elaborated in detail. Finally, the drawbacks of DEP-based systems and the outlook for the future are addressed. This article will aid in advancing technology by bridging the gap between bioscience and engineering. We hope the insights presented in this review will improve cell suspension medium and promote DEP-viable bioparticle manipulation for health-care diagnostics and therapeutics.
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Affiliation(s)
- Revathy Deivasigamani
- Institute of Microengineering and Nanoelectronics (IMEN), Universiti Kebangsaan Malaysia (UKM), Bangi, Selangor, Malaysia
| | - Nur Nasyifa Mohd Maidin
- Institute of Microengineering and Nanoelectronics (IMEN), Universiti Kebangsaan Malaysia (UKM), Bangi, Selangor, Malaysia
| | - Nur Shahira Abdul Nasir
- Institute of Microengineering and Nanoelectronics (IMEN), Universiti Kebangsaan Malaysia (UKM), Bangi, Selangor, Malaysia
| | | | - Aminuddin Bin Ahmad Kayani
- Functional Materials and Microsystems Research Group and the Micro Nano Research Facility, RMIT University, Melbourne, Australia.,ARC Research Hub for Connected Sensors for Health, RMIT University, Melbourne, Australia
| | - Mohd Ambri Mohamed
- Institute of Microengineering and Nanoelectronics (IMEN), Universiti Kebangsaan Malaysia (UKM), Bangi, Selangor, Malaysia
| | - Muhamad Ramdzan Buyong
- Institute of Microengineering and Nanoelectronics (IMEN), Universiti Kebangsaan Malaysia (UKM), Bangi, Selangor, Malaysia
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10
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Hamilton S, Shea D, Ibsen S, Brasino M. On-chip dielectrophoretic recovery and detection of a lactate sensing probiotic from model human saliva. Electrophoresis 2023; 44:442-449. [PMID: 36401837 PMCID: PMC10090127 DOI: 10.1002/elps.202200214] [Citation(s) in RCA: 4] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/26/2022] [Revised: 10/28/2022] [Accepted: 11/06/2022] [Indexed: 11/20/2022]
Abstract
Early detection has led to increased survival for multiple cancers; however, the 5-year survival rate of oral carcinoma (OC) has remained at 40% for the last several decades. Screening for OC is routinely done via visual examinations, followed by tissue biopsy and laboratory testing. Point-of-care testing would be a more convenient and widely available alternative for at-risk individuals. Increased lactate production is a hallmark of many head-and-neck tumors, due to the Warburg Effect, where tumor cells favor glycolysis in the place of oxidative phosphorylation. To detect excess lactate, we have modified the commensal bacterium Escherichia coli Nissle 1917 to express fluorescent reporter genes in response to extracellular lactate. Administering this commensal as a mouth wash and subsequently collecting saliva for the detection of the reporter may allow for noninvasive, early detection of cancerous lesions in at-risk individuals. Furthermore, we demonstrate a new on-chip electrokinetic technique to recover these probiotic probes from model saliva fluid to improve the detection of reporter gene activation.
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Affiliation(s)
- Sean Hamilton
- Cancer Early Detection Advanced Research Center, Knight Cancer Institute, Oregon Health and Science University, Portland, OR 97201, USA
- Department of Biomedical Engineering, School of Medicine, Oregon Health and Science University, Portland, OR 97201, USA
| | - Delaney Shea
- Cancer Early Detection Advanced Research Center, Knight Cancer Institute, Oregon Health and Science University, Portland, OR 97201, USA
- Department of Biomedical Engineering, School of Medicine, Oregon Health and Science University, Portland, OR 97201, USA
| | - Stuart Ibsen
- Cancer Early Detection Advanced Research Center, Knight Cancer Institute, Oregon Health and Science University, Portland, OR 97201, USA
- Department of Biomedical Engineering, School of Medicine, Oregon Health and Science University, Portland, OR 97201, USA
| | - Michael Brasino
- Cancer Early Detection Advanced Research Center, Knight Cancer Institute, Oregon Health and Science University, Portland, OR 97201, USA
- Department of Biomedical Engineering, School of Medicine, Oregon Health and Science University, Portland, OR 97201, USA
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11
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Low‐cost, high‐throughput and rapid‐prototyped 3D‐integrated dielectrophoretic channels for continuous cell enrichment and separation. Electrophoresis 2022. [DOI: 10.1002/elps.202200234] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/12/2022] [Revised: 10/29/2022] [Accepted: 11/06/2022] [Indexed: 11/22/2022]
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12
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Labeed FH, Beale AD, Schneider P, Kitcatt SJ, Kruchek EJ, Reece SE. Circadian rhythmicity in murine blood: Electrical effects of malaria infection and anemia. Front Bioeng Biotechnol 2022; 10:994487. [PMID: 36440448 PMCID: PMC9686327 DOI: 10.3389/fbioe.2022.994487] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/14/2022] [Accepted: 10/27/2022] [Indexed: 11/11/2022] Open
Abstract
Circadian rhythms are biological adaptations to the day-night cycle, whereby cells adapt to changes in the external environment or internal physiology according to the time of day. Whilst many cellular clock mechanisms involve gene expression feedback mechanisms, clocks operate even where gene expression is absent. For example, red blood cells (RBCs) do not have capacity for gene expression, and instead possess an electrophysiological oscillator where cytosolic potassium plays a key role in timekeeping. We examined murine blood under normal conditions as well as in two perturbed states, malaria infection and induced anemia, to assess changes in baseline cellular electrophysiology and its implications for the electrophysiological oscillator. Blood samples were analyzed at 4-h intervals over 2 days by dielectrophoresis, and microscopic determination of parasitemia. We found that cytoplasmic conductivity (indicating the concentration of free ions in the cytoplasm and related to the membrane potential) exhibited circadian rhythmic behavior in all three cases (control, malaria and anemia). Compared to control samples, cytoplasm conductivity was decreased in the anemia group, whilst malaria-infected samples were in antiphase to control. Furthermore, we identified rhythmic behavior in membrane capacitance of malaria infected cells that was not replicated in the other samples. Finally, we reveal the historically famous rhythmicity of malaria parasite replication is in phase with cytoplasm conductivity. Our findings suggest the electrophysiological oscillator can impact on malaria parasite replication and/or is vulnerable to perturbation by rhythmic parasite activities.
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Affiliation(s)
- Fatima H. Labeed
- Centre for Biomedical Engineering, University of Surrey, Guildford, United Kingdom,*Correspondence: Fatima H. Labeed,
| | - Andrew D. Beale
- Centre for Biomedical Engineering, University of Surrey, Guildford, United Kingdom
| | - Petra Schneider
- Institute of Ecology and Evolution, University of Edinburgh, Edinburgh, United Kingdom
| | - Stephen J. Kitcatt
- Centre for Biomedical Engineering, University of Surrey, Guildford, United Kingdom
| | - Emily J. Kruchek
- Centre for Biomedical Engineering, University of Surrey, Guildford, United Kingdom
| | - Sarah E. Reece
- Institute of Ecology and Evolution, University of Edinburgh, Edinburgh, United Kingdom
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13
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Musso N, Romano A, Bonacci PG, Scandura G, Pandino C, Camarda M, Russo GI, Di Raimondo F, Cacciola E, Cacciola R. Label-Free Enrichment of Circulating Tumor Plasma Cells: Future Potential Applications of Dielectrophoresis in Multiple Myeloma. Int J Mol Sci 2022; 23:ijms231912052. [PMID: 36233350 PMCID: PMC9569623 DOI: 10.3390/ijms231912052] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/19/2022] [Revised: 09/23/2022] [Accepted: 09/28/2022] [Indexed: 12/05/2022] Open
Abstract
In multiple myeloma (MM), circulating tumor plasma cells (CTPCs) are an emerging prognostic factor, offering a promising and minimally invasive means for longitudinal patient monitoring. Recent advances highlight the complex biology of plasma cell trafficking, highlighting the phenotypic and genetic signatures of intra- and extra-medullary MM onset, making CTPC enumeration and characterization a new frontier of precision medicine for MM patients, requiring novel technological platforms for their standardized and harmonized detection. Dielectrophoresis (DEP) is an emerging label-free cell manipulation technique to separate cancer cells from healthy cells in peripheral blood samples, based on phenotype and membrane capacitance that could be successfully tested to enumerate and isolate CTPCs. Herein, we summarize preclinical data on DEP development for CTPC detection, as well as their clinical and research potential.
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Affiliation(s)
- Nicolò Musso
- Department of Biomedical and Biotechnological Sciences (BIOMETEC), University of Catania, 95125 Catania, Italy
- StLab SRL, 95126 Catania, Italy
| | - Alessandra Romano
- Department of General Surgery and Surgical Medical Specialties, University of Catania, 95125 Catania, Italy
- Correspondence: ; Tel.: +39-095-378-2971
| | - Paolo Giuseppe Bonacci
- Department of Biomedical and Biotechnological Sciences (BIOMETEC), University of Catania, 95125 Catania, Italy
| | - Grazia Scandura
- Department of General Surgery and Surgical Medical Specialties, University of Catania, 95125 Catania, Italy
| | - Clarissa Pandino
- Department of General Surgery and Surgical Medical Specialties, University of Catania, 95125 Catania, Italy
| | | | - Giorgio Ivan Russo
- Urology Section, University of Catania, Via S. Sofia 78, 95125 Catania, Italy
| | - Francesco Di Raimondo
- Department of General Surgery and Surgical Medical Specialties, University of Catania, 95125 Catania, Italy
| | - Emma Cacciola
- Department of Medical, Surgical Sciences and Advanced Technologies “G.F. Ingrassia”, University of Catania, 95123 Catania, Italy
- Hemostasis/Hematology Unit, A.O.U. Policlinico “G. Rodolico-San Marco”, 95123 Catania, Italy
| | - Rossella Cacciola
- Hemostasis/Hematology Unit, A.O.U. Policlinico “G. Rodolico-San Marco”, 95123 Catania, Italy
- Department of Clinical and Experimental Medicine, University of Catania, 95123 Catania, Italy
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14
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Hughes MP, Fry CH, Labeed FH. Cytoplasmic anion/cation imbalances applied across the membrane capacitance may form a significant component of the resting membrane potential of red blood cells. Sci Rep 2022; 12:15005. [PMID: 36056086 PMCID: PMC9440063 DOI: 10.1038/s41598-022-19316-z] [Citation(s) in RCA: 7] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/17/2022] [Accepted: 08/26/2022] [Indexed: 11/24/2022] Open
Abstract
Electrical aspects of cell function manifest in many ways. The most widely studied is the cell membrane potential, Vm, but others include the conductance and capacitance of the membrane, the conductance of the enclosed cytoplasm, as well as the charge at the cell surface (an electrical double layer) producing an extracellular electrical potential, the ζ-potential. Empirical relationships have been identified between many of these, but not the mechanisms that link them all. Here we examine relationships between Vm and the electrical conductivities of both the cytoplasm and extracellular media, using data from a suspensions of red blood cells. We have identified linear relationships between extracellular medium conductivity, cytoplasm conductivity and Vm. This is in contrast to the standard model of a resting membrane potential which describes a logarithmic relationship between Vm and the concentration of permeable ions in the extracellular medium. The model here suggests that Vm is partially electrostatic in origin, arising from a charge imbalance at an inner electrical double-layer, acting across the membrane and double-layer capacitances to produce a voltage. This model describes an origin for coupling between Vm and ζ, by which cells can alter their electrostatic relationship with their environment, with implications for modulation of membrane ion transport, adhesion of proteins such as antibodies and wider cell-cell interactions.
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Affiliation(s)
- Michael Pycraft Hughes
- Department of Biomedical Engineering, Khalifa University, Abu Dhabi, United Arab Emirates.
- Centre for Biomedical Engineering, University of Surrey, Guildford, GU2 7XH, Surrey, UK.
| | - Christopher H Fry
- School of Physiology, Pharmacology and Neuroscience, University of Bristol, Bristol, BS8 1TD, UK
| | - Fatima H Labeed
- Centre for Biomedical Engineering, University of Surrey, Guildford, GU2 7XH, Surrey, UK
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15
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Hughes MP, Kruchek EJ, Gibbins JM, Griffiths OV, Abdal B, Labeed FH. The Platelet Electrome: Evidence for a Role in Regulation of Function and Surface Interaction. Bioelectricity 2022. [DOI: 10.1089/bioe.2021.0044] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/24/2022] Open
Affiliation(s)
- Michael Pycraft Hughes
- Centre for Biomedical Engineering, University of Surrey, Guildford, United Kingdom
- Department of Biomedical Engineering, Khalifa University, Abu Dhabi, United Arab Emirates
| | - Emily J. Kruchek
- Centre for Biomedical Engineering, University of Surrey, Guildford, United Kingdom
| | - Jonathan M. Gibbins
- Institute for Cardiovascular and Metabolic Research, School of Biological Sciences, University of Reading, Reading, United Kingdom
| | | | - Bader Abdal
- Centre for Biomedical Engineering, University of Surrey, Guildford, United Kingdom
| | - Fatima H. Labeed
- Centre for Biomedical Engineering, University of Surrey, Guildford, United Kingdom
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16
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Wang M, Zhang J, Tan H, Chen D, Lei Y, Li Y, Wang J, Chen J. Inherent Single-Cell Bioelectrical Parameters of Thousands of Neutrophils, Eosinophils and Basophils Derived from Impedance Flow Cytometry. Cytometry A 2022; 101:639-647. [PMID: 35419939 DOI: 10.1002/cyto.a.24559] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/12/2021] [Revised: 03/20/2022] [Accepted: 04/01/2022] [Indexed: 11/08/2022]
Abstract
Single-cell bioelectrical properties are commonly used for blood cell phenotyping in a label-free manner. However, previously reported inherent single-cell bioelectrical parameters (e.g., diameter Dc , specific membrane capacitance Csm and cytoplasmic conductivity σcy ) of neutrophils, eosinophils and basophils were obtained from only tens of individual cells with limited statistical significance. In this study, granulocytes were separated into neutrophils, eosinophils and basophils based on fluorescent flow cytometry, which were further aspirated through a constriction-microchannel impedance flow cytometry for electrical property characterization. Based on this microfluidic impedance flow cytometry, single-cell values of Dc , Csm and σcy were measured as 10.25 ± 0.66 μm, 2.17 ± 0.30 μF/cm2 , and 0.37 ± 0.05 S/m for neutrophils (ncell = 9 442); 9.73 ± 0.51 μm, 2.07 ± 0.19 μF/cm2 , and 0.30 ± 0.04 S/m for eosinophils (ncell = 2 982); 9.75 ± 0.49 μm, 2.06 ± 0.17 μF/cm2 , and 0.31 ± 0.04 S/m for basophils (ncell = 5 377). Based on these inherent single-cell bioelectrical parameters, neural pattern recognition was conducted, producing classification rates of 80.8% (neutrophil vs. eosinophil), 77.7% (neutrophil vs. basophil) and 59.3% (neutrophil vs. basophil). These results indicate that as inherent single-cell bioelectrical parameters, Dc , Csm and σcy can be used to classify neutrophils from eosinophils or basophils to some extent while they cannot be used to effectively distinguish eosinophils from basophils.
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Affiliation(s)
- Minruihong Wang
- State Key Laboratory of Transducer Technology, Aerospace Information Research Institute, Chinese Academy of Sciences, Beijing, People's Republic of China.,School of Future Technology, University of Chinese Academy of Sciences, Beijing, People's Republic of China
| | - Jie Zhang
- CAS Key Laboratory of Genomic and Precision Medicine, Collaborative Innovation Center of Genetics and Development, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, People's Republic of China.,China National Center for Bioinformation, Beijing, People's Republic of China
| | - Huiwen Tan
- State Key Laboratory of Transducer Technology, Aerospace Information Research Institute, Chinese Academy of Sciences, Beijing, People's Republic of China.,School of Future Technology, University of Chinese Academy of Sciences, Beijing, People's Republic of China
| | - Deyong Chen
- State Key Laboratory of Transducer Technology, Aerospace Information Research Institute, Chinese Academy of Sciences, Beijing, People's Republic of China.,School of Future Technology, University of Chinese Academy of Sciences, Beijing, People's Republic of China.,School of Electronic, Electrical and Communication Engineering, University of Chinese Academy of Sciences, Beijing, People's Republic of China
| | - Ying Lei
- CAS Key Laboratory of Genomic and Precision Medicine, Collaborative Innovation Center of Genetics and Development, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, People's Republic of China.,China National Center for Bioinformation, Beijing, People's Republic of China
| | - Yueying Li
- CAS Key Laboratory of Genomic and Precision Medicine, Collaborative Innovation Center of Genetics and Development, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, People's Republic of China.,China National Center for Bioinformation, Beijing, People's Republic of China
| | - Junbo Wang
- State Key Laboratory of Transducer Technology, Aerospace Information Research Institute, Chinese Academy of Sciences, Beijing, People's Republic of China.,School of Future Technology, University of Chinese Academy of Sciences, Beijing, People's Republic of China.,School of Electronic, Electrical and Communication Engineering, University of Chinese Academy of Sciences, Beijing, People's Republic of China
| | - Jian Chen
- State Key Laboratory of Transducer Technology, Aerospace Information Research Institute, Chinese Academy of Sciences, Beijing, People's Republic of China.,School of Future Technology, University of Chinese Academy of Sciences, Beijing, People's Republic of China.,School of Electronic, Electrical and Communication Engineering, University of Chinese Academy of Sciences, Beijing, People's Republic of China
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17
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Yang G, Wei L, Thong BKS, Fu Y, Cheong IH, Kozlakidis Z, Li X, Wang H, Li X. A Systematic Review of Oral Biopsies, Sample Types, and Detection Techniques Applied in Relation to Oral Cancer Detection. BIOTECH 2022; 11:5. [PMID: 35822813 PMCID: PMC9245907 DOI: 10.3390/biotech11010005] [Citation(s) in RCA: 14] [Impact Index Per Article: 4.7] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/18/2022] [Revised: 02/18/2022] [Accepted: 02/24/2022] [Indexed: 12/11/2022] Open
Abstract
Background: Early identification of the stage of oral cancer development can lead to better treatment outcomes and avoid malignant transformation. Therefore, this review aims to provide a comprehensive overview that describes the development of standardized procedures for oral sample collection, characterization, and molecular risk assessment. This can help investigators to choose the appropriate sampling method and downstream analyses for different purposes. Methods: This systematic review was conducted according to the PRISMA guidelines. Using both PubMed and Web of Science databases, four independent authors conducted a literature search between 15 and 21 June 2021. We used key search terms to broaden the search for studies. Non-conforming articles were removed using an EndNote-based and manual approach. Reviewers used a designed form to extract data. Results: This review included a total of 3574 records, after eliminating duplicate articles and excluding papers that did not meet the inclusion criteria. Finally, 202 articles were included in this review. We summarized the sampling methods, biopsy samples, and downstream analysis. The biopsy techniques were classified into tissue and liquid biopsy. The common sequential analysis of tissue biopsy includes histopathological examination such as H&E or IHC to identify various pathogenic features. Meanwhile, liquid samples such as saliva, blood, and urine are analyzed for the purpose of screening to detect mutations in cancer. Commonly used technologies are PCR, RT-PCR, high-throughput sequencing, and metabolomic analysis. Conclusions: Currently, tissue biopsies provide increased diagnostic value compared to liquid biopsy. However, the minimal invasiveness and convenience of liquid biopsy make it a suitable method for mass screening and eventual clinical adoption. The analysis of samples includes histological and molecular analysis. Metabolite analysis is rising but remains scarce.
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Affiliation(s)
- Guanghuan Yang
- State Key Laboratory of Oncogenes and Related Genes, Centre for Single-Cell Omics, School of Public Health, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China; (G.Y.); (L.W.); (B.K.S.T.); (Y.F.); (I.H.C.); (X.L.)
| | - Luqi Wei
- State Key Laboratory of Oncogenes and Related Genes, Centre for Single-Cell Omics, School of Public Health, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China; (G.Y.); (L.W.); (B.K.S.T.); (Y.F.); (I.H.C.); (X.L.)
| | - Benjamin K. S. Thong
- State Key Laboratory of Oncogenes and Related Genes, Centre for Single-Cell Omics, School of Public Health, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China; (G.Y.); (L.W.); (B.K.S.T.); (Y.F.); (I.H.C.); (X.L.)
| | - Yuanyuan Fu
- State Key Laboratory of Oncogenes and Related Genes, Centre for Single-Cell Omics, School of Public Health, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China; (G.Y.); (L.W.); (B.K.S.T.); (Y.F.); (I.H.C.); (X.L.)
| | - Io Hong Cheong
- State Key Laboratory of Oncogenes and Related Genes, Centre for Single-Cell Omics, School of Public Health, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China; (G.Y.); (L.W.); (B.K.S.T.); (Y.F.); (I.H.C.); (X.L.)
| | - Zisis Kozlakidis
- International Agency for Research on Cancer, World Health Organization, 69372 Lyon, France;
| | - Xue Li
- State Key Laboratory of Oncogenes and Related Genes, Centre for Single-Cell Omics, School of Public Health, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China; (G.Y.); (L.W.); (B.K.S.T.); (Y.F.); (I.H.C.); (X.L.)
| | - Hui Wang
- State Key Laboratory of Oncogenes and Related Genes, Centre for Single-Cell Omics, School of Public Health, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China; (G.Y.); (L.W.); (B.K.S.T.); (Y.F.); (I.H.C.); (X.L.)
| | - Xiaoguang Li
- State Key Laboratory of Oncogenes and Related Genes, Centre for Single-Cell Omics, School of Public Health, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China; (G.Y.); (L.W.); (B.K.S.T.); (Y.F.); (I.H.C.); (X.L.)
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18
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Tan H, Wang M, Zhang Y, Huang X, Chen D, Li Y, Wu MH, Wang K, Wang J, Chen J. Inherent Bioelectrical Parameters of Hundreds of Thousands of Single Leukocytes Based on Impedance Flow Cytometry. Cytometry A 2022; 101:630-638. [PMID: 35150049 DOI: 10.1002/cyto.a.24544] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/17/2021] [Revised: 01/15/2022] [Accepted: 02/02/2022] [Indexed: 11/11/2022]
Abstract
As label-free biomarkers, bioelectrical properties of single cells have been widely used in hematology analyzers for 3-part differential of leukocytes, in which, however, instrument dependent bioelectrical parameters (e.g., DC/AC impedance values) rather than inherent bioelectrical parameters (e.g., diameter Dc , specific membrane capacitance Csm and cytoplasmic conductivity σcy ) were used, leading to poor comparisons among different instruments. In order to address this issue, this study collected inherent bioelectrical parameters from hundreds of thousands of white blood cells based on a home-developed impedance flow cytometry with corresponding 3-part differential of leukocytes realized. More specifically, leukocytes were separated into three major subtypes of granulocytes, monocytes and lymphocytes based on density gradient centrifugation. Then these separated cells were aspirated through a constriction-microchannel based impedance flow cytometry where inherent bioelectrical parameters of Dc , Csm and σcy were quantified as 9.8 ± 0.7 μm, 2.06 ± 0.26 μF/cm2 , and 0.34 ± 0.05 S/m for granulocytes (ncell = 134 829); 10.4 ± 1.0 μm, 2.45 ± 0.48 μF/cm2 , and 0.42 ± 0.08 S/m for monocytes (ncell = 40 226); 8.0 ± 0.5 μm, 2.23 ± 0.34 μF/cm2 , and 0.35 ± 0.08 S/m for lymphocytes (ncell = 129 193). Based on these inherent bioelectrical parameters, neural pattern recognition was conducted, producing a high "classification accuracy" of 93.5% in classifying these three subtypes of leukocytes. These results indicate that as inherent bioelectrical parameters, Dc , Csm and σcy can be used to electrically phenotype white blood cells in a label-free manner.
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Affiliation(s)
- Huiwen Tan
- State Key Laboratory of Transducer Technology, Aerospace Information Research Institute, Chinese Academy of Sciences, Beijing, People's Republic of China.,School of Future Technology, University of Chinese Academy of Sciences, Beijing, People's Republic of China
| | - Minruihong Wang
- State Key Laboratory of Transducer Technology, Aerospace Information Research Institute, Chinese Academy of Sciences, Beijing, People's Republic of China.,School of Future Technology, University of Chinese Academy of Sciences, Beijing, People's Republic of China
| | - Yi Zhang
- State Key Laboratory of Transducer Technology, Aerospace Information Research Institute, Chinese Academy of Sciences, Beijing, People's Republic of China.,School of Electronic, Electrical and Communication Engineering, University of Chinese Academy of Sciences, Beijing, People's Republic of China
| | - Xukun Huang
- State Key Laboratory of Transducer Technology, Aerospace Information Research Institute, Chinese Academy of Sciences, Beijing, People's Republic of China.,School of Electronic, Electrical and Communication Engineering, University of Chinese Academy of Sciences, Beijing, People's Republic of China
| | - Deyong Chen
- State Key Laboratory of Transducer Technology, Aerospace Information Research Institute, Chinese Academy of Sciences, Beijing, People's Republic of China.,School of Future Technology, University of Chinese Academy of Sciences, Beijing, People's Republic of China.,School of Electronic, Electrical and Communication Engineering, University of Chinese Academy of Sciences, Beijing, People's Republic of China
| | - Yueying Li
- CAS Key Laboratory of Genomic and Precision Medicine, Collaborative Innovation Center of Genetics and Development, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, People's Republic of China.,China National Center for Bioinformation, Beijing, People's Republic of China
| | - Min-Hsien Wu
- Graduate Institute of Biochemical and Biomedical Engineering, Chang Gung University, Taoyuan City, Taiwan, Republic of China
| | - Ke Wang
- School of Electronic Engineering, Beijing University of Posts and Telecommunications, Beijing, People's Republic of China
| | - Junbo Wang
- State Key Laboratory of Transducer Technology, Aerospace Information Research Institute, Chinese Academy of Sciences, Beijing, People's Republic of China.,School of Future Technology, University of Chinese Academy of Sciences, Beijing, People's Republic of China.,School of Electronic, Electrical and Communication Engineering, University of Chinese Academy of Sciences, Beijing, People's Republic of China
| | - Jian Chen
- State Key Laboratory of Transducer Technology, Aerospace Information Research Institute, Chinese Academy of Sciences, Beijing, People's Republic of China.,School of Future Technology, University of Chinese Academy of Sciences, Beijing, People's Republic of China.,School of Electronic, Electrical and Communication Engineering, University of Chinese Academy of Sciences, Beijing, People's Republic of China
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19
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Duncan JL, Davalos RV. A review: Dielectrophoresis for characterizing and separating similar cell subpopulations based on bioelectric property changes due to disease progression and therapy assessment. Electrophoresis 2021; 42:2423-2444. [PMID: 34609740 DOI: 10.1002/elps.202100135] [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] [Received: 05/06/2021] [Revised: 09/19/2021] [Accepted: 09/23/2021] [Indexed: 12/16/2022]
Abstract
This paper reviews the use of dielectrophoresis for high-fidelity separations and characterizations of subpopulations to highlight the recent advances in the electrokinetic field as well as provide insight into its progress toward commercialization. The role of cell subpopulations in heterogeneous clinical samples has been studied to deduce their role in disease progression and therapy resistance for instances such as cancer, tissue regeneration, and bacterial infection. Dielectrophoresis (DEP), a label-free electrokinetic technique, has been used to characterize and separate target subpopulations from mixed samples to determine disease severity, cell stemness, and drug efficacy. Despite its high sensitivity to characterize similar or related cells based on their differing bioelectric signatures, DEP has been slowly adopted both commercially and clinically. This review addresses the use of dielectrophoresis for the identification of target cell subtypes in stem cells, cancer cells, blood cells, and bacterial cells dependent on cell state and therapy exposure and addresses commercialization efforts in light of its sensitivity and future perspectives of the technology, both commercially and academically.
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Affiliation(s)
- Josie L Duncan
- Bioelectromechanical Systems Laboratory, Department of Mechanical Engineering, Virginia Tech, Blacksburg, Virginia, USA.,Bioelectromechanical Systems Laboratory, Wake Forest School of Biomedical Engineering and Sciences, Virginia Tech, Blacksburg, Virginia, USA
| | - Rafael V Davalos
- Bioelectromechanical Systems Laboratory, Department of Mechanical Engineering, Virginia Tech, Blacksburg, Virginia, USA.,Bioelectromechanical Systems Laboratory, Wake Forest School of Biomedical Engineering and Sciences, Virginia Tech, Blacksburg, Virginia, USA
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20
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Dielectrophoresis as a Tool to Reveal the Potential Role of Ion Channels and Early Electrophysiological Changes in Osteoarthritis. MICROMACHINES 2021; 12:mi12080949. [PMID: 34442571 PMCID: PMC8402151 DOI: 10.3390/mi12080949] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 07/13/2021] [Revised: 07/30/2021] [Accepted: 08/08/2021] [Indexed: 11/16/2022]
Abstract
Diseases such as osteoarthritis (OA) are commonly characterized at the molecular scale by gene expression and subsequent protein production; likewise, the effects of pharmaceutical interventions are typically characterized by the effects of molecular interactions. However, these phenomena are usually preceded by numerous precursor steps, many of which involve significant ion influx or efflux. As a consequence, rapid assessment of cell electrophysiology could play a significant role in unravelling the mechanisms underlying drug interactions and progression of diseases, such as OA. In this study, we used dielectrophoresis (DEP), a technique that allows rapid, label-free determination of the dielectric parameters to assess the role of potassium ions on the dielectric characteristics of chondrocytes, and to investigate the electrophysiological differences between healthy chondrocytes and those from an in vitro arthritic disease model. Our results showed that DEP was able to detect a significant decrease in membrane conductance (6191 ± 738 vs. 8571 ± 1010 S/m2), membrane capacitance (10.3 ± 1.47 vs. 14.5 ± 0.01 mF/m2), and whole cell capacitance (5.4 ± 0.7 vs. 7.5 ± 0.3 pF) following inhibition of potassium channels using 10 mM tetraethyl ammonium, compared to untreated healthy chondrocytes. Moreover, cells from the OA model had a different response to DEP force in comparison to healthy cells; this was seen in terms of both a decreased membrane conductivity (782 S/m2 vs. 1139 S/m2) and a higher whole cell capacitance (9.58 ± 3.4 vs. 3.7 ± 1.3 pF). The results show that DEP offers a high throughput method, capable of detecting changes in membrane electrophysiological properties and differences between disease states.
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21
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Henslee EA. Review: Dielectrophoresis in cell characterization. Electrophoresis 2020; 41:1915-1930. [DOI: 10.1002/elps.202000034] [Citation(s) in RCA: 17] [Impact Index Per Article: 3.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/02/2020] [Revised: 05/31/2020] [Accepted: 07/14/2020] [Indexed: 12/23/2022]
Affiliation(s)
- Erin A. Henslee
- Department of Engineering Wake Forest University 455 Vine St. Winston‐Salem USA
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22
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Hoque R, Mostafid H, Hughes MP. Rapid, Low-Cost Dielectrophoretic Diagnosis of Bladder Cancer in a Clinical Setting. IEEE JOURNAL OF TRANSLATIONAL ENGINEERING IN HEALTH AND MEDICINE 2020; 8:4300405. [PMID: 32656002 PMCID: PMC7343103 DOI: 10.1109/jtehm.2020.3004743] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 04/11/2020] [Revised: 05/28/2020] [Accepted: 06/19/2020] [Indexed: 11/10/2022]
Abstract
Bladder cancer is the 9th most common cancer worldwide. Diagnosing bladder cancer typically involves highly invasive cystoscopy, with followup monitored using uteroscopy. Molecular methods have been developed as an adjunct to this, but tend to be expensive or require expert operator input. Here we present a study of the use of dielectrophoresis (DEP) of voided cells from eight cancer-presenting patients and eight healthy controls as an alternative low-cost and operator-independent method of bladder cancer detection. This study suggests that there are statistically significant differences ([Formula: see text]) between characteristics of the DEP spectrum of clinical samples, and that using this marker we were able to obtain sensitivity of 75% and specificity of 88%, in line with many molecular methods; exclusion of samples where a DEP spectrum is not present (due to low cell counts) increased sensitivity to 100%, showing this can be improved by increasing the cell collection rate. As samples were analyzed a day after collection, we suggest that the method may be amenable to a centralized mail-in analysis service.
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Affiliation(s)
- Rashedul Hoque
- Centre for Biomedical Engineering, School of Mechanical Engineering SciencesUniversity of SurreyGuildfordGU2 7XHU.K.
| | | | - Michael Pycraft Hughes
- Centre for Biomedical Engineering, School of Mechanical Engineering SciencesUniversity of SurreyGuildfordGU2 7XHU.K.
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23
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Jing H, Yang X, Wu S, Zhang M, Zhou J, Yuan J, Zhu Z, Huang K. A numerical coupling method for particle tracking in electromagnetic fields. THE EUROPEAN PHYSICAL JOURNAL. E, SOFT MATTER 2019; 42:48. [PMID: 31011841 DOI: 10.1140/epje/i2019-11810-3] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 01/24/2019] [Accepted: 03/11/2019] [Indexed: 06/09/2023]
Abstract
With the arrival of the information age, the electromagnetic energy in space increases constantly, resulting in the influence of electromagnetic waves on the charged aerosol particles in the environment which should be taken into account. Here, a numerical coupling method based on temporal and spatial scales is proposed to solve the difficulty in obtaining the trajectory of particles under the action of high-frequency electromagnetic waves. In the temporal scale, two constant forces with linear relationship are used to equilibrate the electromagnetic field forces under different conditions, however the above-mentioned equivalent method has the space limitation; in addition, on the spatial scale, the model with larger geometry should be divided into multiple basic modules spatially, the domain division method is adopted and due to the above method it can be used well in the basic module. Verified the correctness through the comparison of the results, and compared with the traditional method, the above method greatly reduces the computational complexity. Some interesting results were obtained by calculating the modulated waves with the above method, which indicate that special forms of electromagnetic waves will significantly affect the motion of particles.
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Affiliation(s)
- Heng Jing
- College of Electronics and Information Engineering, Sichuan University, 610065, Chengdu, China
| | - Xiaoqing Yang
- College of Electronics and Information Engineering, Sichuan University, 610065, Chengdu, China.
| | - Shiyue Wu
- College of Electronics and Information Engineering, Sichuan University, 610065, Chengdu, China
| | - Man Zhang
- College of Electronics and Information Engineering, Sichuan University, 610065, Chengdu, China
| | - Jie Zhou
- College of Electronics and Information Engineering, Sichuan University, 610065, Chengdu, China
| | - Jianping Yuan
- School of Astronautics, Northwest Polytechnical University, 710072, Xi'an, China
| | - Zhanxia Zhu
- School of Astronautics, Northwest Polytechnical University, 710072, Xi'an, China
| | - Kama Huang
- College of Electronics and Information Engineering, Sichuan University, 610065, Chengdu, China
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24
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Nerguizian V, Stiharu I, Al-Azzam N, Yassine-Diab B, Alazzam A. The effect of dielectrophoresis on living cells: crossover frequencies and deregulation in gene expression. Analyst 2019; 144:3853-3860. [DOI: 10.1039/c9an00320g] [Citation(s) in RCA: 25] [Impact Index Per Article: 4.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/09/2023]
Abstract
This article documents the effect of dielectrophoresis on living cells.
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Affiliation(s)
- Vahé Nerguizian
- Department of Electrical Engineering
- École de technologie supérieure (ÉTS)
- Montreal
- Canada
| | - Ion Stiharu
- Department of Mechanical and Industrial Engineering
- Concordia University
- Montreal
- Canada
| | - Nosayba Al-Azzam
- Department of Physiology and Biochemistry
- Jordan University of Science and Technology
- Irbid
- Jordan
| | | | - Anas Alazzam
- Department of Mechanical Engineering
- Khalifa University
- Abu Dhabi
- United Arab Emirates
- Department of Electrical Engineering
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25
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Wang K, Chang CC, Chiu TK, Zhao X, Chen D, Chou WP, Zhao Y, Wang HM, Wang J, Wu MH, Chen J. Membrane capacitance of thousands of single white blood cells. J R Soc Interface 2018; 14:rsif.2017.0717. [PMID: 29212758 DOI: 10.1098/rsif.2017.0717] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/30/2017] [Accepted: 11/01/2017] [Indexed: 12/18/2022] Open
Abstract
As label-free biomarkers, the electrical properties of single cells are widely used for cell type classification and cellular status evaluation. However, as intrinsic cellular electrical markers, previously reported membrane capacitances (e.g. specific membrane capacitance Cspec and total membrane capacitance Cmem) of white blood cells were derived from tens of single cells, lacking statistical significance due to low cell numbers. In this study, white blood cells were first separated into granulocytes and lymphocytes by density gradient centrifugation and were then aspirated through a microfluidic constriction channel to characterize both Cspec and Cmem Thousands of granulocytes (ncell = 3327) and lymphocytes (ncell = 3302) from 10 healthy blood donors were characterized, resulting in Cspec values of 1.95 ± 0.22 µF cm-2 versus 2.39 ± 0.39 µF cm-2 and Cmem values of 6.81 ± 1.09 pF versus 4.63 ± 0.57 pF. Statistically significant differences between granulocytes and lymphocytes were located for both Cspec and Cmem In addition, neural network-based pattern recognition was used to classify white blood cells, producing successful classification rates of 78.1% for Cspec and 91.3% for Cmem, respectively. These results indicate that as intrinsic bioelectrical markers, membrane capacitances may contribute to the classification of white blood cells.
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Affiliation(s)
- Ke Wang
- State Key Laboratory of Transducer Technology, Institute of Electronics, Chinese Academy of Sciences, Beijing, People's Republic of China.,University of Chinese Academy of Sciences, Beijing, People's Republic of China
| | - Chun-Chieh Chang
- Graduate Institute of Biochemical and Biomedical Engineering, Chang Gung University, Taoyuan City, Taiwan, Republic of China
| | - Tzu-Keng Chiu
- Department of Chemical and Materials Engineering, Chang Gung University, Taoyuan City, Taiwan, Republic of China
| | - Xiaoting Zhao
- Department of Cellular and Molecular Biology, Beijing Chest Hospital, Capital Medical University, Beijing, People's Republic of China
| | - Deyong Chen
- State Key Laboratory of Transducer Technology, Institute of Electronics, Chinese Academy of Sciences, Beijing, People's Republic of China.,University of Chinese Academy of Sciences, Beijing, People's Republic of China
| | - Wen-Pin Chou
- Graduate Institute of Biochemical and Biomedical Engineering, Chang Gung University, Taoyuan City, Taiwan, Republic of China
| | - Yang Zhao
- Institute of Microelectronics, Chinese Academy of Sciences, Beijing, People's Republic of China
| | - Hung-Ming Wang
- Division of Haematology/Oncology, Department of Internal Medicine, Chang Gung Memorial Hospital at Linkou, Taoyuan City, Taiwan, Republic of China
| | - Junbo Wang
- State Key Laboratory of Transducer Technology, Institute of Electronics, Chinese Academy of Sciences, Beijing, People's Republic of China .,University of Chinese Academy of Sciences, Beijing, People's Republic of China
| | - Min-Hsien Wu
- Graduate Institute of Biochemical and Biomedical Engineering, Chang Gung University, Taoyuan City, Taiwan, Republic of China .,Division of Haematology/Oncology, Department of Internal Medicine, Chang Gung Memorial Hospital at Linkou, Taoyuan City, Taiwan, Republic of China
| | - Jian Chen
- State Key Laboratory of Transducer Technology, Institute of Electronics, Chinese Academy of Sciences, Beijing, People's Republic of China .,University of Chinese Academy of Sciences, Beijing, People's Republic of China
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Mahabadi S, Labeed FH, Hughes MP. Dielectrophoretic analysis of treated cancer cells for rapid assessment of treatment efficacy. Electrophoresis 2018; 39:1104-1110. [PMID: 29405335 DOI: 10.1002/elps.201700488] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/18/2017] [Revised: 01/26/2018] [Accepted: 01/30/2018] [Indexed: 12/21/2022]
Abstract
Whilst personalized medicine (where interventions are precisely tailored to a patient's genotype and phenotype, as well as the nature and state of the disease) is regarded as an optimal form of treatment, the time and cost associated with it means it remains inaccessible to the greater public. A simpler alternative, stratified medicine, identifies groups of patients who are likely to respond to a given treatment. This allows appropriate treatments to be selected at the start of therapy, avoiding the common "trial and error" approach of replacing a therapy only once it is demonstrated to be ineffective in the patient. For stratification to be effective, tests are required that rapidly predict treatment effectiveness. Most tests use genetic analysis to identify drug targets, but these can be expensive and may not detect changes in the phenotype that affect drug sensitivity. An alternative method is to assess the whole-cell phenotype by evaluating drug response using cells from a biopsy. We assessed dielectrophoresis to assess drug efficacy on short timescales and at low cost. To explore the principle of assessing drug efficacy we examined two cell lines (one expressing EGFR, one not) with the drug Iressa. We then further explored the sensitive cells using combinations of chemotherapeutic and radiotherapeutic therapies. Our results compare with known effects of these cell/treatment combination, and offer the additional benefit over methods such as TUNEL of detecting drug effects such as cell cycle arrest, which do not cause cell death.
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Affiliation(s)
- Sina Mahabadi
- Centre for Biomedical Engineering, Department of Mechanical Engineering Sciences, University of Surrey, Guildford, UK
| | - Fatima H Labeed
- Centre for Biomedical Engineering, Department of Mechanical Engineering Sciences, University of Surrey, Guildford, UK
| | - Michael P Hughes
- Centre for Biomedical Engineering, Department of Mechanical Engineering Sciences, University of Surrey, Guildford, UK
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Abd Rahman N, Ibrahim F, Yafouz B. Dielectrophoresis for Biomedical Sciences Applications: A Review. SENSORS 2017; 17:s17030449. [PMID: 28245552 PMCID: PMC5375735 DOI: 10.3390/s17030449] [Citation(s) in RCA: 120] [Impact Index Per Article: 15.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 10/26/2016] [Revised: 12/10/2016] [Accepted: 12/20/2016] [Indexed: 12/18/2022]
Abstract
Dielectrophoresis (DEP) is a label-free, accurate, fast, low-cost diagnostic technique that uses the principles of polarization and the motion of bioparticles in applied electric fields. This technique has been proven to be beneficial in various fields, including environmental research, polymer research, biosensors, microfluidics, medicine and diagnostics. Biomedical science research is one of the major research areas that could potentially benefit from DEP technology for diverse applications. Nevertheless, many medical science research investigations have yet to benefit from the possibilities offered by DEP. This paper critically reviews the fundamentals, recent progress, current challenges, future directions and potential applications of research investigations in the medical sciences utilizing DEP technique. This review will also act as a guide and reference for medical researchers and scientists to explore and utilize the DEP technique in their research fields.
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Affiliation(s)
- Nurhaslina Abd Rahman
- Department of Biomedical Engineering, Faculty of Engineering, University of Malaya, 50603 Kuala Lumpur, Malaysia.
- Centre for Innovation in Medical Engineering (CIME), Faculty of Engineering, University of Malaya, 50603 Kuala Lumpur, Malaysia.
| | - Fatimah Ibrahim
- Department of Biomedical Engineering, Faculty of Engineering, University of Malaya, 50603 Kuala Lumpur, Malaysia.
- Centre for Innovation in Medical Engineering (CIME), Faculty of Engineering, University of Malaya, 50603 Kuala Lumpur, Malaysia.
| | - Bashar Yafouz
- Department of Biomedical Engineering, Faculty of Engineering, University of Malaya, 50603 Kuala Lumpur, Malaysia.
- Centre for Innovation in Medical Engineering (CIME), Faculty of Engineering, University of Malaya, 50603 Kuala Lumpur, Malaysia.
- Faculty of Engineering and Information Technology, Taiz University, 6803 Taiz, Yemen.
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28
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Matsuoka H. ELECTROCHEMISTRY 2016; 84:41-49. [DOI: 10.5796/electrochemistry.84.41] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/05/2022] Open
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