title: Laser Capture Microdissection of Archival Kidney Tissue for qRT-PCR.
abstract: Whole-organ molecular analysis of the kidney potentially misses important factors involved in the pathogenesis of disease in glomeruli and tubules. Organ wide analysis can however be augmented by using laser capture microdissection (LCM) to isolate morphologically similar cells and nephron structures from a heterogeneous tissue section via direct visualization of the cells. The protocol here provides a practical approach utilizing LCM in combination with RNA isolation techniques for downstream analysis. This technique is readily applicable to study mRNA expression in isolated glomeruli and tubules in both experimental animal models and human kidney biopsy material.
authors.affiliation: Department of Nephrology, The Royal Melbourne Hospital, 300 Grattan Street, Parkville, Melbourne, VIC, 3050, Australia. tim.hewitson@mh.org.au.|Department of Medicine, University of Melbourne, Melbourne, VIC, 3050, Australia. tim.hewitson@mh.org.au.
authors.forename: Tim D
authors.initials: TD
authors.lastname: Hewitson
authors.affiliation: Department of Anatomical Pathology, The Royal Melbourne Hospital, 300 Grattan Street, Parkville, Melbourne, VIC, 3050, Australia.
authors.forename: Michael
authors.initials: M
authors.lastname: Christie
authors.affiliation: Department of Nephrology, The Royal Melbourne Hospital, 300 Grattan Street, Parkville, Melbourne, VIC, 3050, Australia.
authors.forename: Edward R
authors.initials: ER
authors.lastname: Smith
mesh_terms: D000818:Animals; D006801:Humans; D007668:Kidney / Q000166:cytology* / Q000378:metabolism*; D007678:Kidney Glomerulus / Q000166:cytology / Q000378:metabolism; D007684:Kidney Tubules / Q000166:cytology / Q000378:metabolism; D061346:Laser Capture Microdissection / Q000379:methods; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods
keywords: Glomeruli; Kidney; Laser capture microdissection; PCR; RNA; Tubules