Microsurgery and microinjection techniques in mitosis research

Charles A. Day, Jessica Hornick, Alyssa Langfald, Christopher Mader, Edward H. Hinchcliffe

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

The use of microtechnique for studying cell division is well established (Begg & Ellis, 1979; Wadsworth, 1999; Zhang & Nicklas, 1999). The advantage of microinjection in cell division research is the timed delivery of a macromolecules at a particular stage of mitosis (for example, pre- vs postanaphase), which can circumvent the spindle assembly checkpoint (Hinchcliffe et al., 2016). Micromanipulation can be used to remove whole organelles, such as the centrosome or nucleus and examine the effects on cell division (Hinchcliffe et al., 2001; Hornick et al., 2011). The focus of this chapter is on methods for microinjection and micromanipulation of cultured mammalian cells. We describe pulling and shaping microneedles, as well as the imaging chambers we use. We also provide information on cell culture conditions, and imaging techniques used for our long-term observation studies, which allow cells to be followed on the order of several days.

Original languageEnglish (US)
Title of host publicationMethods in Cell Biology
EditorsHelder Maiato, Melina Schuh
PublisherAcademic Press Inc.
Pages159-172
Number of pages14
ISBN (Print)9780128141427
DOIs
StatePublished - 2018

Publication series

NameMethods in Cell Biology
Volume145
ISSN (Print)0091-679X

Bibliographical note

Funding Information:
We would like to thank Mr. Rick Miller and Prof. Kip Sluder for introducing us to the world of microtechnique and for their foundational contributions to the use of live-cell microscopy for the study of cell cycle regulation and mitosis. Work in the author's lab supported by funds from the American Cancer Society, the Hormel Foundation, the 5th District Eagles Cancer Telethon, Austin MN's “Paint the Town Pink”, the US Department of Defense (CDMRP), and the National Institutes of Health.

Publisher Copyright:
© 2018 Elsevier Inc.

Copyright:
Copyright 2018 Elsevier B.V., All rights reserved.

Keywords

  • Antibody
  • Cell cycle
  • Checkpoint
  • Long-term time-lapse
  • Microneedle

PubMed: MeSH publication types

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

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