6533b827fe1ef96bd1285951
RESEARCH PRODUCT
Cell Proliferation High-Content Screening on Adherent Cell Cultures
José PertusaJosé Manuel Morante-redolatPau Carrillo-barberàsubject
0303 health sciences030219 obstetrics & reproductive medicineCell growthChemistryCell cycleMolecular biologyNeural stem cellDeoxyuridine03 medical and health scienceschemistry.chemical_compound0302 clinical medicineLabellingHigh-content screeningImmunostainingEx vivo030304 developmental biologydescription
Pulse-chase experiments using 5-bromo-2'-deoxyuridine (BrdU), or the more recent EdU (5-etynil-2'-deoxyuridine), enable the identification of cells going through S phase. This chapter describes a high-content proliferation assay pipeline for adherent cell cultures. High-throughput imaging is followed by high-content data analysis using a non-supervised ImageJ macroinstruction that segments the individual nuclei, determines the nucleoside analogue absence/presence, and measures the signal of up to two additional nuclear markers. Based upon the specific combination with proliferation-specific protein immunostaining, the percentage of cells undergoing different phases of the cell cycle (G0, G1, S, G2, and M) might be established. The method can be also used to estimate the proliferation (S phase) rate of particular cell subpopulations identified through labelling with specific nuclear markers.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2019-01-01 |