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Fixation of immune cells


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#1 tatzilo

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Posted 03 August 2009 - 06:58 AM

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 :)

#2 cellcounter

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Posted 03 August 2009 - 08:48 AM

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 :(


You can try out cytospin or frozen sections of agar-embedded block. See protocols here.

#3 little mouse

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Posted 03 August 2009 - 10:18 PM

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.




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