CCD microscopy and image analysis of cells and chromosomes stained by fluorescence in situ hybridization

Summary This paper reviews methods and applications of CCD microscopy for analysing cells and chromosomes subjected to fluorescence in situ hybridization (FISH). The current status of indirect and direct FISH staining methods with respect to probe labelling, detection sensitivity, multiplicity and DNA resolution is summarized. Microscope hardware, including special multi-band pass filters and CCD cameras required for FISH analysis, is described. Then follows a detailed discussion of current and emerging applications such as the automated enumeration of chromosomal abnormalities (counting of dots in interphase cells), comparative genomic hybridization, automated evaluation of radiation-induced chromosomal translocations, and high-resolution DNA mapping on highly extended chromatin. Finally, the limitations of the present methodology and future prospects are discussed.The Historical Journal lecture for 1993 presented by Dr Tanke to a joint meeting of the Royal Microscopical society of cell Biology, Clinical Cytology, Electron Microscopy and Pathology and the Belgian Socities in Brussels, Belgium on 2 September 1993.This work was supported financially by the Netherlands Organization for Scientific Research (NWO) and in part by Imagenetics, Framingham, MA, USA.