Laser-controlled microdissection of tissues opens a window of new opportunities.
Gene expression analysis using total RNA of bulk tissue usually cannot assign specific messages to particular cell types. Cell-specific RNA expression profiling, though, may be crucial for a better understanding ofthe role of each distinct cell type within a physiological or pathophysiological setting. RNA profiling based on laser-controlled microdissection (LCM) of defined cells of a tissue now provides a useful tool for studying molecular crosstalk between different cell types within a tissue. The LCM technique allows for efficient isolation of single cells with no or very low contamination of surrounding tissue components, simultaneously leaving the intracellular structure and molecules intact. In this review, different issues of the LCM technique and the RNA amplification procedure for microarray analysis are discussed. An exemplary summary of results obtained from gene profiling of epithelial and stromal cells from human prostate tumors is presented, demonstrating the power of LCM-based molecular analysis. Finally, we discuss the potential use of the LCM technique i) to study the transcriptome of distinct cells from formalin-fixed and paraffin-embedded tissues in subcellular RNA profiling and ii) high resolution proteomic and metabolistic studies.
Hergenhahn
Manfred
M
Division C040: Genetic Alterations in Carcinogenesis, German Cancer Research Center, Heidelberg, Germany.
Kenzelmann
Marc
M
Gröne
Hermann-Josef
HJ
D004210:Dissection / Q000295:instrumentation / Q000379:methods; D020869:Gene Expression Profiling / Q000379:methods; D006801:Humans; D007834:Lasers; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D012333:RNA, Messenger / Q000032:analysis*
D012333:RNA, Messenger