Immunohistochemical staining, laser capture microdissection, and filter‐aided sample preparation‐assisted proteomic analysis of target cell populations within tissue samples. Issue 11 (27th April 2013)

• Record Type: Journal Article
• Authors: He, Jintang; Zhu, Jianhui; Liu, Yashu; Wu, Jing; Nie, Song; Heth, Jason A.; Muraszko, Karin M.; Fan, Xing; Lubman, David M.
• Is Part Of: Electrophoresis. Volume 34:Issue 11(2013:Jun.)
• Page Start: 1627
• Page End: 1636
• Publication Date: 2013-04-27
• Subjects: Immunohistochemical staining -- Laser capture microdissection -- Proteomics -- Target cell population -- Tissue sample; Electrophoresis -- Periodicals; Electrophoresis -- Periodicals; 541.372
• DOI: 10.1002/elps.201200566
• Languages: English
• ISSNs: 0173-0835

Abstract

Abstract : An important problem involves isolating subpopulations of cells defined by protein markers in clinical tissue samples for proteomic studies. We describe a method termedI mmunohistochemical staining, laser capture microdissection (L CM) and filter‐aided sample preparation (F ASP)‐A ssistedP roteomic analysis ofT arget cell populations within tissue samples (ILFAPT). The principle of ILFAPT is that a target cell population expressing a protein of interest can be lit up by immunohistochemical staining and isolated from tissue sections using LCM for FASP and proteomic analysis. Using this method, we isolated a small population of CD90 + stem‐like cells from glioblastoma multiforme tissue sections and identified 674 high‐confidence (false discovery rate < 0.01) proteins from 32 nL of CD90 + cells by LC‐MS/MS using an Orbitrap Elite mass spectrometer. We further quantified the relative abundance of proteins identified from equal volumes of LCM‐captured CD90 + and CD90 − cells, where 109 differentially expressed proteins were identified. The major group of these differentially expressed proteins was relevant to cell adhesion and cellular movement. This ILFAPT method has demonstrated the ability to provide in‐depth proteome analysis of a very small specific cell population within tissues. It can be broadly applied to the study of target cell populations within clinical specimens.