Manipulating cultured mammalian cells for mitosis research

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish (US)
Title of host publicationMethods in Cell Biology
EditorsPhong Tran
PublisherAcademic Press Inc.
Pages43-61
Number of pages19
Volume158
ISBN (Print)9780128200087
DOIs
StatePublished - 2020

Publication series

NameMethods in cell biology
PublisherAcademic Press Inc.
ISSN (Print)0091-679X

Bibliographical note

Funding Information:
Work in the author's lab is supported by grants from NIH (HL125353) and the DoD CDMRP (CA171071 and CA130436), and support from the I.J. Holton Trust and the Hormel Foundation.

Publisher Copyright:
© 2020 Elsevier Inc.

Keywords

  • Cell culture
  • Cell cycle
  • Chilling
  • Chromosomes
  • Cold-dependent
  • Microinjection
  • Microscopy
  • Microtubule
  • Spindle
  • Spinning disk

PubMed: MeSH publication types

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

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