Good Day:
I am looking for a protocol for fixation of PBMC's and MSC's (Human) that will allow for surface analysis by flow cytometry.
Thx and best regards...dwf
Many of the CD antigens (CD3, CD4, etc) are ruined by fixation with aldehyde fixatives. You can fix with ethanol or methanol and that will preserve those antigens. I don't recall the percent of ethanol; it might have been 95% and used it cold (2-8C). After 5 minutes, wash to get the cells back into saline or PBS for staining.
I have stained the cells always on ice, and therefore have used 2% fresh (!) prepared Formaldehyde at 4ºC for 5-10 minutes, then wash 2 times with icecold buffer, ...
Hello Daniel, Are you fixing to mitigate potential biological hazard or transport of unlabelled tissue? I fix with freshly(
- Badges
- Science method