Cremer Marion, Müller Stefan, Köhler Daniela, Brero Alessandro, Solovei Irina
Ludwig-Maximilians University Munich, Department of Biology II, AG Thomas Cremer (Chair of Anthropology and Human Genetics), 82152 Martinsried-Planegg, Germany.
CSH Protoc. 2007 May 1;2007:pdb.prot4723. doi: 10.1101/pdb.prot4723.
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.
引言
三维保存细胞核的荧光原位杂交（3D-FISH）结合三维显微镜和图像重建，是在单细胞水平上分析不同核靶点（如整个染色体区域、染色体亚区域或单基因座）排列的有效工具。本方案重点介绍培养的哺乳动物细胞的固定、预处理和3D-FISH。它可应用于多种贴壁或悬浮生长的细胞类型。
