Basic Research
Copyright ©The Author(s) 2003. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jun 15, 2003; 9(6): 1337-1341
Published online Jun 15, 2003. doi: 10.3748/wjg.v9.i6.1337
Gene-expression analysis of single cells-nested polymerase chain reaction after laser microdissection
Xin Shi, JÖrg Kleeff, Zhao-Wen Zhu, Bruno Schmied, Wen-Hao Tang, Arthur Zimmermann, Markus W. BÜchler, Helmut Friess
Xin Shi, JÖrg Kleeff, Zhao-Wen Zhu, Bruno Schmied, Markus W. BÜchler, Helmut Friess, Department of General Surgery, University of Heidelberg, Im Neuenheimer Feld 110, D-69120 Heidelberg, Germany
Xin Shi, Wen-Hao Tang, Department of General Surgery, Zhongda Hospital, Southeast University, Nanjing 210009, Jiangsu Province, P. R. China
Arthur Zimmermann, Institute of Pathology, University of Bern, Inselspital, CH-3010 Bern, Switzerland
Author contributions: All authors contributed equally to the work.
Correspondence to: Helmut Friess, M.D.Department of General Surgery, University of Heidelberg, Im Neuenheimer Feld 110, D-69120 Heidelberg, Germany. helmut_friess@
Telephone: +49-6221-566900 Fax: +49-6221-566903
Received: December 30, 2002
Revised: March 4, 2003
Accepted: March 12, 2003
Published online: June 15, 2003

AIM: The structural and functional characteristics of cells are dependent on the specific gene expression profile. The ability to study and compare gene expression at the cellular level will therefore provide valuable insights into cell physiology and pathophysiology.

METHODS: Individual cells were isolated from frozen colon tissue sections using laser microdissection. DNA as well as RNA were extracted, and total RNA was reversely transcribed to complementary DNA (cDNA). Both DNA and cDNA were analyzed by nested polymerase chain reaction (PCR). The quality of isolated DNA and RNA was satisfactory.

RESULTS: Single cells were successfully microdissected using an ultraviolet laser micromanipulator. Nested PCR amplification products of DNA and cDNA of single cells could clearly be visualized by agarose gel electrophoresis.

CONCLUSION: The combined use of laser microdissection and nested-PCR provides an opportunity to analyze gene expression in single cells. This method allows the analysis and identification of specific genes which are involved in physiological and pathophysiological processes in a complex of variable cell phenotypes.

Keywords: $[Keywords]