Quantitative Proteomic Analysis of Mass Limited Tissue Samples for Spatially Resolved Tissue Profiling

Methods Mol Biol. 2018;1788:269-277. doi: 10.1007/7651_2017_78.

Abstract

Traditionally, proteomic studies have been carried out on whole tissues or organs enabling the profiling of thousands of proteins within a single LC-MS analysis. A disadvantage of this approach is that proteomes generated from whole tissues are an "average" that represents a blend of cell types and distinct anatomical regions which can obscure important biological phenomena. Laser capture microdissection (LCM) is an elegant method that allows tissue features of interest, as small as a single cell, to be identified and isolated for downstream analysis. Herein we describe an approach that utilizes an immobilized enzyme reactor (IMER) coupled directly to nanoLC-MS/MS for highly sensitive, automated, quantitative proteomic analysis of the microscopic tissue specimens generated by LCM.

Keywords: Immobilized enzyme reactor; Laser capture microdissection; Mass spectrometry; NanoLC; Nanoproteomics; Proteomics.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Enzymes, Immobilized / chemistry
  • Equipment Design
  • Humans
  • Laser Capture Microdissection / instrumentation*
  • Laser Capture Microdissection / methods
  • Proteome / analysis*
  • Proteomics / instrumentation
  • Proteomics / methods*
  • Sample Size
  • Solid Phase Extraction / instrumentation
  • Solid Phase Extraction / methods
  • Tandem Mass Spectrometry / instrumentation
  • Tandem Mass Spectrometry / methods*

Substances

  • Enzymes, Immobilized
  • Proteome