Fixation of immune cells - "how to" protocol needed (Aug/03/2009 )

Hi,

up to now we found a lot of ideas how to fix suspension cells onto a surface. The best seems to be working with 4% PFA.
But when it comes to carry out the experiment a few questions arose:

- centrifugation or waiting to settle down the cells
- how to apply PFA not to bring the cells into suspension

Thanks in advance :)

tatzilo on Aug 3 2009, 06:58 AM said:

Hi,

up to now we found a lot of ideas how to fix suspension cells onto a surface. The best seems to be working with 4% PFA.
But when it comes to carry out the experiment a few questions arose:

- centrifugation or waiting to settle down the cells
- how to apply PFA not to bring the cells into suspension

Thanks in advance

If you don't have a cytospin, you can let the cells settle down on a polylysin coated glass slide. wash, do your labeling and fix. I did it with CD8+ T cells.