Cite as: Cold Spring Harb. Protoc.; 2007; doi:10.1101/pdb.prot4723

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Cell Preparation and Multicolor FISH in 3D Preserved Cultured Mammalian Cells

Marion Cremer1, Stefan Müller, Daniela Köhler, Alessandro Brero, and Irina Solovei

Ludwig-Maximilians University Munich, Department of Biology II, AG Thomas Cremer (Chair of Anthropology and Human Genetics), 82152 Martinsried-Planegg, Germany

1Corresponding author (Marion.Cremer@lrz.uni-muenchen.de)


INTRODUCTION

Fluorescence 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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