Microsurgery and microinjection techniques in mitosis research

• Published in: Methods in cell biology Vol. 145; pp. 159 - 172
• Main Authors: Day, Charles A, Hornick, Jessica, Langfald, Alyssa, Mader, Christopher, Hinchcliffe, Edward H
• Format: Book Chapter Journal Article
• Language: English
• Published: United States Elsevier Science & Technology 2018
• Subjects: Animals; Centrosome - physiology; Humans; Microinjections - methods; Micromanipulation - methods; Microsurgery - methods; Mitosis - physiology; Spindle Apparatus - physiology; Checkpoint; Antibody; Long-term time-lapse; Microneedle; Cell cycle
• ISBN: 9780128141427; 0128141425
• ISSN: 0091-679X
• DOI: 10.1016/bs.mcb.2018.03.020

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.