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Help! methanol fixation on SH-SY5Y cells - immunofluorescence (Oct/10/2006 )

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I am using methanol to fix SH-SY5Y cells, and stain nuclear with DAPI afterwards. the condition i am using is: ice-cold methanol fix for 5 min. I found DAPI stain sometimes goes out of the nuclear and looks fluffy. I have no idea what is wrong!!
Please, please help me, thanks.

-moon-

we use formaldehyde or PFA for fixing cells (SHSY5Y). coud u try one of these ?

-scolix-

QUOTE (scolix @ Oct 11 2006, 08:15 AM)
we use formaldehyde or PFA for fixing cells (SHSY5Y). coud u try one of these ?

I used that, that's fine. Just the antibody I am using specificly like methanol fixation. Did you try methanol fixation on SH-SY5y? THanks

-moon-

QUOTE (moon @ Oct 12 2006, 01:38 PM)
I used that, that's fine. Just the antibody I am using specificly like methanol fixation. Did you try methanol fixation on SH-SY5y? THanks


unfortunately not.

-scolix-

maybe you can try Carnoy's fixative(1 part glacial acetic acid to 3 parts methanol).

-vivien-