Abstract
Although several microscopic techniques are available for separation of small groups of cells from surrounding tissue, the high energy concentrated into a small area, the easy control of beam position and the lack of direct contact with the material to be dissected make lasers the best option for a large scale microdissection. Laser microdissection technology provides samples of homogenous cells, or even a single cell, isolated from whole tissue or cytological materials to ensure that biological molecules such as DNA, RNA or protein are undamaged during sampling in order to define the molecular and cellular biology of diseases, including cancer. This article reviews the various techniques of laser microdissection including capture, catapulting and gravity-assisted, in addition to a cheaper alternative (ultrasound). Methods of sample preparation, including contamination eradication, enrichment, fixation, staining and extraction of biological molecules are also reviewed. Application of genomic and proteomic technology to laser microdissected cells has revolutionized the classification of solid cancer and enhanced its diagnosis, prognosis and drug discovery.
Footnotes
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Abbreviations: CCD, charged coupled device; cDNA, copy deoxyribonucleic acid; DSP, dithio-bis(succinimidyl propionate); EASI, epithelial aggregate separation and isolation; FFT, fresh frozen tissue; FFPET, formalin fixed and paraffin embedded tissue; GAM, gravity-assisted microdissection; H&E, hematoxylin and eosin; IR, infrared; LCM, laser capture microdissection; LMD, laser microdissection; LPC, laser pressure catapulting; mRNA, messenger ribonucleic acid; NA, numerical aperture; PEN, polyethylene naphthalate; PET, polyethylene tetraphthalate; POL, polyester; RT-PCR, reverse transcription polymerase chain reaction; mRNA, messenger ribonucleic acid;UV, ultraviolet light.
- Received May 12, 2006.
- Accepted May 23, 2006.
- Copyright© 2006 International Institute of Anticaner Research (Dr. John G. Delinassios), All rights reserved