INTRODUCTIONFluorescence in situ hybridization (FISH) on three-dimensional preserved nuclei (3D-FISH) in combination with three-dimensional-microscopy and image reconstruction is an efficient tool to analyze the arrangement of distinct nuclear targets such as entire chromosome territories, chromosomal subregions, or single gene loci on a single-cell level. This protocol focuses on fixation, pretreatments, and 3D-FISH on cultured mammalian cells. It can be applied to a variety of cell types growing adherently or in suspension.
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