| Chapter title |
Cell Cycle Analysis Using In Vivo Staining of DNA-Synthesizing Cells.
|
|---|---|
| Chapter number | 228 |
| Book title |
Imaging and Tracking Stem Cells
|
| Published in |
Methods in molecular biology, May 2019
|
| DOI | 10.1007/7651_2019_228 |
| Pubmed ID | |
| Book ISBNs |
978-1-07-160626-1, 978-1-07-160627-8
|
| Authors |
Páral, Petr, Báječný, Martin, Savvulidi, Filipp, Nečas, Emanuel, Petr Páral, Martin Báječný, Filipp Savvulidi, Emanuel Nečas |
| Abstract |
The thymidine analogues BrdU (5-bromo-2´-deoxyuridine) and EdU (5-ethynyl-2´-deoxyuridine) are routinely used for determination of the cells synthesizing DNA in the S-phase of the cell cycle. Availability of the anti-BrdU antibody clone MoBu-1 detecting only BrdU allowed to develop a method for the sequential DNA labelling by these two thymidine analogues for determining the cell cycle kinetic parameters.In the current step-by-step protocol, we present` two approaches optimized for in vivo study of the cell cycle and the limitations that such approaches imply: (1) determination of the cell flow rate into the G2-phase by dual EdU/BrdU DNA-labelling method and (2) determination of the outflow of DNA-labelled cells arising from the mitosis. |
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