Dynamic proliferation assessment in flow cytometry.
Curr Protoc Cell Biol
; Chapter 8: Unit 8.6.1-23, 2010 Sep.
Article
em En
| MEDLINE
| ID: mdl-20853345
ABSTRACT
Dynamic proliferation assessment via flow cytometry is legitimately supposed to be the most powerful tool for recording cell cycle kinetics in-vitro. The preeminent feature is a single cell-based multi-informative analysis by temporal high-resolution. Flow cytometric approaches are based on labeling of proliferating cells via thymidine substitution by a base analog (e.g., 5-bromo-2'-deoxyuridine, BrdU) that is added to cell cultures either for a short period of time (pulse labeling) or continuously until cell harvesting. This unit describes the alternative use of the thymidine analog 5-ethynyl-2'-deoxyuridine (EdU) in place of BrdU for three different applications (1) dynamic proliferation assessment by EdU pulse cell labeling; (2) the same approach as (1) but in combination with live/dead cell discrimination; and (3) dynamic cell cycle analysis based on continuous cell labeling with EdU and Hoechst fluorochrome quenching. In contrast to the detection of BrdU incorporation, EdU-positive cells can be identified by taking advantage of click chemistry, which facilitates a simplified and fast cell preparation. Further analysis options but also limitations of the utilization of EdU are discussed.
Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
Ciclo Celular
/
Desoxiuridina
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Proliferação de Células
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Citometria de Fluxo
Limite:
Animals
/
Humans
Idioma:
En
Revista:
Curr Protoc Cell Biol
Ano de publicação:
2010
Tipo de documento:
Article
País de afiliação:
Alemanha