Decordier Ilse, Kirsch-Volders Micheline
Laboratorium voor Cellulaire Genetica, Vrije Universiteit Brussel, Brussels, Belgium.
Methods Mol Biol. 2013;1044:237-44. doi: 10.1007/978-1-62703-529-3_12.
In recent years, cytogenetics in combination with molecular methods has made rapid progress, resulting in new molecular cytogenetic methodologies such as fluorescence in situ hybridization (FISH). FISH is a molecular cytogenetic technique used for the detection of specific chromosomal rearrangements and applicable to many different specimen types. It uses fluorescently labeled DNA probes complementary to regions of individual chromosomes. These labeled DNA segments hybridize with the cytological targets in the sample and can be visualized by fluorescence microscopy in interphase nuclei or on metaphase chromosomes. Here, we describe the FISH methodology with centromeric probes for human cells, which is used in combination with the cytokinesis-block micronucleus assay and which allows discrimination between mutagens inducing DNA breakage (clastogens) or chromosome loss (aneugens).
近年来,细胞遗传学与分子方法相结合取得了快速进展,催生了诸如荧光原位杂交(FISH)等新的分子细胞遗传学方法。FISH是一种分子细胞遗传学技术,用于检测特定的染色体重排,适用于多种不同的样本类型。它使用与单个染色体区域互补的荧光标记DNA探针。这些标记的DNA片段与样本中的细胞学靶标杂交,并可通过荧光显微镜在间期核或中期染色体上观察到。在此,我们描述了用于人类细胞的着丝粒探针FISH方法,该方法与胞质分裂阻断微核试验联合使用,可区分诱导DNA断裂的诱变剂(断裂剂)或染色体丢失的诱变剂(非整倍体剂)。