Materials

Cells of interest

DAPI/sulforhodamine 101/detergent solution (see recipe)

Flow cytometer equipped with UV excitation and filters for collection of blue and red fluorescence

1. Collect ~106 cells from the culture and centrifuge 5 min at 300 x g, room temperature.

2. Suspend the cell pellet in 1 ml DAPI/sulforhodamine 101/detergent solution and vortex 20 sec.

3. Analyze cells on the flow cytometer, using UV excitation (e.g., 351-nm line from an argon-ion laser, or mercury lamp with a UG1 filter). Collect blue DAPI fluorescence in a band from 460 to 500 nm and red fluorescence of sulforhodamine 101 above 600 nm.

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