Application of Laser-Assisted Microdissection for Tissue and Cell-Specific Analysis of RNA, Proteins, and Metabolites

Abstract

The present state of different laser-assisted microdissection methods and their application in various disciplines of plant science is reviewed. The first part of the present review describes the basic effects of a laser beam on biological tissue, and following sections dealing with practical aspects of sample preparation and the technologies implemented in laser-assisted microdissection procedures. An attempt is made to distinguish laser-capture microdissection from laser cutting, to distinguish different varieties of laser cutting, and to discuss the advantages and drawbacks of the various methods for specific applications. The second part of the review covers applications of laser-assisted microdissection in various areas of plant science. Using lasers to conduct microsurgery on plant tissue and to dissect chromosome parts are two highly specialized areas that are discussed. RNA isolation and cell-specific gene expression analysis are the most frequent reasons for using laser-assisted microdissection methods, so a section comprising recent applications is included. Although proteomic methods for analyzing the contents of specific cell populations are available, proteins have been analyzed infrequently in laser-microdissected plant tissue. Therefore, the corresponding section also includes examples from animal samples. The final section is dedicated to the emerging field of cell type-specific metabolite profiling, including secondary metabolites, in extracts of laser-microdissected plant samples by mass spectrometry and nuclear magnetic resonance spectroscopy.