Methods in cellular and molecular pathology

Abstract

This is an overview and introduction to cellular and molecular methods, which are applied in diagnostic and investigative endocrine pathology. In the first part of the chapter, the principles of immunohistochemical techniques are discussed, including necessary controls and an approach to test new antibodies. The second part of the chapter deals with molecular methods. After a general introduction to molecular biology, an overview of methods is provided which are applied in situ to identify DNA, specific mRNAs and chromosomal structures at a cellular level, such as in situ hybridization (ISH), fluorescent in situ hybridization (FISH) and in situ proteomics. The third part of the chapter focuses on liquid-based molecular methods, which are applied in endocrine pathology. They include microdissection of cell groups for molecular analyses, polymerase chain reaction (PCR)-based methods for DNA and RNA detection as well as the analysis of mutations, loss of heterozygosity and clonality. Furthermore, comparative genomic hybridization (CGH) and the RNA expression array technology to identify chromosomal gains and losses as well as gene expression are discussed. © 2010 Springer-Verlag New York.