Protocols >
Formaldehyde Fixation of Cells
Safety Note: Work in hood when dealing with formaldehyde.
- Grow a 2-5ml overnight culture in your media at 30oC with shaking.
- Subculture in fresh media and grow for 4 hrs at 30oC with shaking.
- **work in hood** Add 10%, volume of the culture, of 37% formaldehyde and incubate cultures at 30oC with shaking for 10 min.
- Transfer 1ml of culture to a microcentrifuge tube, in hood, and spin down at 6000 RPM for 3 min.
- Remove supernatant in hood and dispose in formaldehyde waste. Resuspend cells in 900 µl 1X PBS and 100 µl 37% formaldehyde. (for sick cultures, I have spun down an entire 5 ml culture and resuspended in 900 µl 1X PBS and 100 µl 37% formaldehyde)
- Incubate cells at room temp for 1 hr.
- Store cells at 4oC.
