Journal
METHODS
Volume 41, Issue 2, Pages 143-150Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2006.07.022
Keywords
cell cycle analysis; DNA analogues; BrdU; synchronization; mitotic shake-off
Funding
- NCRR NIH HHS [P20 RR15578] Funding Source: Medline
- NIA NIH HHS [R01 AG016694] Funding Source: Medline
- NIGMS NIH HHS [R01 GM41690, R01 GM041690] Funding Source: Medline
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Fluorescence activated cell sorting (FACS) analysis has become a standard tool to analyze cell cycle distributions in populations of cells. These methods require relatively large numbers of cells, and do not provide optimal resolution of the transitions between cell cycle phases. In this report we describe in detail complementary methods that utilize the incorporation of nucleotide analogs combined with microscopic examination. While often more time consuming, these protocols typically require far fewer cells, and allow accurate kinetic assessment of cell cycle progression. We also describe the use of a technique for the synchronization of adherent cells in mitosis by simple mechanical agitation (mitotic shake-off) that eliminates physiological perturbation associated with drug treatments. (c) 2006 Elsevier Inc. All rights reserved.
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