用于有丝分裂研究的培养哺乳动物细胞的操作。
Manipulating cultured mammalian cells for mitosis research.
机构信息
Hormel Institute and Masonic Cancer Center, University of Minnesota, Austin, MN, United States.
Hormel Institute and Masonic Cancer Center, University of Minnesota, Austin, MN, United States.
出版信息
Methods Cell Biol. 2020;158:43-61. doi: 10.1016/bs.mcb.2020.02.001. Epub 2020 Mar 12.
The study of mitosis has always relied on bulk-preparation biochemistry techniques (Mazia & Dan, 1952), but very early on lent itself to living, single cell microscopic techniques (Inoue, 1953; Taylor, 1959). Here we describe several of the methods used by our lab to study cell division in living cultured cells, including cold-induced mitotic arrest, cold-induced chromosome missegregation, same-cell live and fixed cell imaging, and microinjection of inactivating antibodies. We detail our imaging system based on an upright fluorescent microscope and spinning disk confocal, as well as the customized "HEKS" metal support slide imaging chambers.
有丝分裂的研究一直依赖于批量制备的生物化学技术（Mazia 和 Dan，1952），但很早就适合于活的单细胞显微镜技术（Inoue，1953；Taylor，1959）。在这里，我们描述了我们实验室用于研究活培养细胞中细胞分裂的几种方法，包括冷诱导有丝分裂阻滞、冷诱导染色体错误分离、同一细胞的活细胞和固定细胞成像，以及失活抗体的微注射。我们详细介绍了我们基于正置荧光显微镜和旋转盘共聚焦的成像系统，以及定制的“HEKS”金属支撑载玻片成像室。
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