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 language||English (US)|
|Title of host publication||Methods in Cell Biology|
|Editors||Helder Maiato, Melina Schuh|
|Publisher||Academic Press Inc.|
|Number of pages||14|
|State||Published - 2018|
|Name||Methods in Cell Biology|
Bibliographical noteFunding 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.
© 2018 Elsevier Inc.
- Cell cycle
- Long-term time-lapse
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.