HomeLive Go cell cycle analysis with Hoechst 33342 and Pyronin Y
Flow cytometry may be used to determine the level of senescence in a cell population. Go and G1 can be separated by use of Pyronin Y (Sigma) which preferentially binds to RNA and Hoechst 33342 (Sigma) which binds to A-T base pairs. Cells are loaded with Hoechst 33342 at 10 ug/ml and Pyronin Y at 0.5 ug/ml by incubating for 45 mins at 37C. Pyronin Y is excited at 488 nm and emits at 575 nm, while Hoechst 33342 is excited UV lasers (350-360 nm), near UV line (375 nm) or violet diode (405 nm).
