Manipulating cultured mammalian cells for mitosis research

Charles A Day; Alyssa Langfald; Edward H Hinchcliffe

doi:10.1016/bs.mcb.2020.02.001

Manipulating cultured mammalian cells for mitosis research

Methods Cell Biol. 2020:158:43-61. doi: 10.1016/bs.mcb.2020.02.001. Epub 2020 Mar 12.

Authors

Charles A Day¹, Alyssa Langfald¹, Edward H Hinchcliffe²

Affiliations

¹ Hormel Institute and Masonic Cancer Center, University of Minnesota, Austin, MN, United States.
² Hormel Institute and Masonic Cancer Center, University of Minnesota, Austin, MN, United States. Electronic address: hinc0002@umn.edu.

Abstract

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.

Keywords: Cell culture; Cell cycle; Chilling; Chromosomes; Cold-dependent; Microinjection; Microscopy; Microtubule; Spindle; Spinning disk.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Culture Techniques / methods*
Cell Line
Chlorocebus aethiops
Humans
Imaging, Three-Dimensional
Mammals / physiology*
Microinjections
Mitosis*
Rats
Research*
Spindle Apparatus / metabolism

Abstract

Publication types

MeSH terms

Grants and funding