help with fixing suspension cell for staining

Tim Fitzmaurice tjf11 at cus.cam.ac.uk
Wed Jul 23 04:03:31 EST 1997


On Fri, 11 Jul 1997, Changhong Dai wrote:
> Hi, I am interested in methods for fixing suspension cell(say U937) for
> lacZ staining(how to make the suspension cell stick to the culture dish
> or glass for staining). Would anyone kindly share any ideas about how to
> do it?

Well, when I am fixing neurospheres sometimes they float around beforehand
or are just barely resting on the coverslip...if I wash with buffer, fix
for 2 mins in Methanol at -20, wash and then fix with 4% paraformaldehyde
for 5 mins, they tend to stick pretty well after the methanol treatment,
and better afterr the paraformaldehyde.
You might want to try that treatment perhaps after using a cytospin to
put the cells on slides, though I have no idea how the methanol is going
to affect the lacZ staining system.

Tim





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