Transcript analysis of laser microdissected plant cells

Abstract

Laser microdissection (LM) of biological specimens is enabling high‐throughput molecular analysis to be carried out with a cellular or tissue‐specific resolution. If a particular structure is identifiable using a conventional light microscope then it is most likely available for isolation by LM, this includes subcellular structures such as organelles or even individual chromosomes. To obtain reliable data from laser microdissected samples several protocols need to be established and in many cases optimized in a target‐dependent manner. Here we consider technical aspects of plant‐based studies that have used LM for transcript analysis. We provide detail on aspects of tissue preparation, microdissection, RNA yield and quality assessment and describe how RNA amplification can affect the analysis. We discuss protocols from our own lab alongside other user‐developed approaches and highlight new technologies that may prove useful to plant biologists considering the use of LM in their research.