Laser-capture microdissection and pressure catapulting for the analysis of gene expression in the renal glomerulus.
Investigation into the molecular mechanisms regulating normal renal physiology and pathophysiology has benefited from the development of microdissection techniques enabling sampling of specific cell populations or structures within the kidney. Laser-capture microdissection and pressure catapulting is a relatively new, entirely non-contact microdissection technique that facilitates the assay of mRNA and protein expression in single nephron segments or populations. Herein, we describe methods for sample preparation, microdissection and collection of glomeruli from archival renal biopsies for later analysis of gene expression using real-time PCR. Microdissection of glomeruli from archival renal biopsy sections was carried out using the PALM Microbeam UV laser system from P.A.L.M. Technologies.
Edgley
Amanda J
AJ
Department of Medicine, St. Vincent's Hospital, University of Melbourne, Melbourne, VIC, Australia.
Gow
Renae M
RM
Kelly
Darren J
DJ
D000818:Animals; D006651:Histocytochemistry / Q000295:instrumentation / Q000379:methods; D006801:Humans; D007678:Kidney Glomerulus / Q000378:metabolism*; D007834:Lasers; D042282:Microdissection / Q000295:instrumentation* / Q000379:methods*; D011312:Pressure