Laser capture microdissection.
Laser capture microdissection (LCM) offers a rapid and precise method of isolating and removing specified cells from complex tissues for subsequent analysis of their RNA, DNA, or protein content, thereby allowing assessment of the role of the cell type in the normal physiologic or disease process being studied. In this unit, protocols for the preparation of mammalian frozen tissues, fixed tissues, and cytologic specimens for LCM, including hematoxylin and eosin staining, are presented, as well as a protocol for the performance of LCM utilizing the PixCell I or II Laser Capture Microdissection System manufactured by Arcturus Engineering. Also provided is a protocol for tissue processing and paraffin embedding, and recipes for lysis buffers for the recovery of nucleic acids and proteins. The Commentary section addresses the types of specimens that can be utilized for LCM and approaches to staining of specimens for cell visualization. Emphasis is placed on the preparation of tissue or cytologic specimens as this is critical to effective LCM.
Frost
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AR
University of Alabama at Birmingham, Birmingham, Alabama, USA.
Eltoum
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Siegal
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GP
D000818:Animals; D002498:Centrifugation / Q000379:methods; D015925:Cryopreservation / Q000379:methods; D005629:Frozen Sections; D016591:Histocytological Preparation Techniques; D006801:Humans; D007834:Lasers; D008322:Mammals; D042282:Microdissection / Q000295:instrumentation / Q000379:methods*; D016612:Paraffin Embedding / Q000379:methods; D011506:Proteins / Q000032:analysis; D012313:RNA / Q000032:analysis; D013048:Specimen Handling / Q000379:methods; D016707:Tissue Fixation / Q000379:methods
D011506:Proteins; D012313:RNA