Molecular analysis of complex tissues is facilitated by laser capture microdissection: critical role of upstream tissue processing.
Every tissue contains heterogeneous cell populations. Laser capture microdissection (LCM) facilitates cell isolation from complex tissues followed by molecular analysis. LCM entails placing a transparent film over a tissue section or a cytological sample, visualizing the cells microscopically, and selectively adhering the cells of interest to the film with a focused pulse from an infrared laser. The film with the procured cells is then removed from the original sample and placed directly into DNA, RNA, or protein-extraction buffer for processing. LCM has revolutionized molecular analysis of complex tissues because it combines the topographic precision of microscopy with the power of molecular genetics, genomics, and proteomics. However, the success of molecular analysis still depends on the experimental design and requires the understanding of each technical step involved in specimen preparation. This review attempts to rationalize and demystify the choice of various technical options in upstream tissue processing supporting global analytical strategies.
Rekhter
M D
MD
Department of Cardiovascular Pharmacology, Pfizer Global Research and Development, Ann Arbor Laboratories, MI 48105, USA. mark.rekhter@pfizer.com
Chen
J
J
D000818:Animals; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D004247:DNA / Q000032:analysis; D004210:Dissection / Q000295:instrumentation / Q000379:methods*; D005821:Genetic Techniques; D006801:Humans; D007150:Immunohistochemistry; D008853:Microscopy / Q000295:instrumentation / Q000379:methods*; D012313:RNA / Q000032:analysis
D012313:RNA; D004247:DNA