Hi there,
we are trying to establish stable transfected NIH 3T3 or C2C12 cells by FACS sorting (on plastic dishes), and starting off with 90 % green cells after the sort, we end up with hardly any by one week or so afterwards. Even using pEGFP-C2 as a control construct. Also, we do not see massive cell death.
Any suggesting what the cause might be? In the end, these are commonly used cell lines, and I did not expect this kind of problem ...
Thanks
Do untransfected cells survive, when you are running them through the FACS? Are there any preservatives (sodium azide?) in the sheath fluid?
Depending on the size of the cells you might want to sort based on a larger droplet size to minimize stress to the cells (i.e. 100um nozzle instead of 70um). This increases droplet size but mostly helps in reducing the pressure (i.e. 40 PSI to 15 PSI).
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line