Bizard Anna H, Nielsen Christian F, Hickson Ian D
Department of Cellular and Molecular Medicine, Center for Chromosome Stability, University of Copenhagen, Panum Institute, 2200, Copenhagen, N, Denmark.
Methods Mol Biol. 2018;1672:495-508. doi: 10.1007/978-1-4939-7306-4_33.
Ultrafine anaphase bridges (UFBs) are thin DNA threads linking the separating sister chromatids in the anaphase of mitosis. UFBs are thought to form when topological DNA entanglements between two chromatids are not resolved prior to anaphase onset. In contrast to other markers of defective chromosome segregation, UFBs cannot be detected by direct staining of the DNA, but instead can be detected using immunofluorescence-based approaches. Due to the fact that they are short-lived and fragile in nature, UFBs can be challenging to detect. In this chapter, we describe methods that have been optimized for successful detection of UFBs. We also provide guidelines for the optimization of UFBs detection depending on the antibody and the cell line to be used.
超微后期桥(UFBs)是在有丝分裂后期连接分离的姐妹染色单体的细DNA丝。UFBs被认为是在两个染色单体之间的拓扑DNA缠结在后期开始前未得到解决时形成的。与染色体分离缺陷的其他标志物不同,UFBs不能通过DNA的直接染色来检测,而是可以使用基于免疫荧光的方法来检测。由于它们本质上寿命短且脆弱,UFBs的检测可能具有挑战性。在本章中,我们描述了为成功检测UFBs而优化的方法。我们还提供了根据所使用的抗体和细胞系优化UFBs检测的指南。