Koehler Daniela, Zakhartchenko Valeri, Ketterl Nina, Wolf Eckhard, Cremer Thomas, Brero Alessandro
Institute of Human Genetics, Biozentrum (LMU), University of Münich, Planegg-Martinsried, Germany.
Methods Mol Biol. 2010;659:437-45. doi: 10.1007/978-1-60761-789-1_34.
Fluorescence in situ hybridization (FISH) is a commonly used technique for the visualization of whole chromosomes or subchromosomal regions, such as chromosome arms, bands, centromeres, or single gene loci. FISH is routinely performed on chromosome spreads, as well as on three-dimensionally preserved cells or tissues (3D FISH). We have developed 3D FISH protocol for mammalian preimplantation embryos to investigate the nuclear organization of chromosome territories and subchromosomal regions during the first developmental stages. In contrast to cells, embryos have much more depth and their nuclei are therefore less accessible to probes used to visualize specific genomic regions by FISH. The present protocol was developed to establish a balance between sufficient embryo permeabilization and maximum preservation of nuclear morphology.
荧光原位杂交（FISH）是一种常用技术，用于观察整条染色体或亚染色体区域，如染色体臂、带、着丝粒或单基因座。FISH通常在染色体涂片上进行，也在三维保存的细胞或组织上进行（三维FISH）。我们已经开发出用于哺乳动物植入前胚胎的三维FISH方案，以研究发育早期阶段染色体区域和亚染色体区域的核组织。与细胞不同，胚胎有更大的深度，因此用于通过FISH观察特定基因组区域的探针更难进入其细胞核。本方案旨在实现足够的胚胎通透化与最大程度保持核形态之间的平衡。
