Edgley Amanda J, Gow Renae M, Kelly Darren J
Department of Medicine, St. Vincent's Hospital, University of Melbourne, Melbourne, VIC, Australia.
Methods Mol Biol. 2010;611:29-40. doi: 10.1007/978-1-60327-345-9_3.
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.
对调节正常肾脏生理和病理生理的分子机制的研究受益于显微切割技术的发展，该技术能够对肾脏内特定细胞群或结构进行采样。激光捕获显微切割和压力弹射是一种相对较新的、完全非接触式的显微切割技术，有助于分析单个肾单位节段或细胞群中的mRNA和蛋白质表达。在此，我们描述了从存档肾活检组织中制备样本、显微切割和收集肾小球的方法，以便随后使用实时PCR分析基因表达。使用P.A.L.M. Technologies公司的PALM Microbeam紫外激光系统对存档肾活检切片中的肾小球进行显微切割。
