Methanol Fixation of SH-SY5Y cells for Immunofluorescence

wexfrdo · #1 01-28-2009, 03:14 AM

Hi,
Im using methanol to fix SH-SY5Y cells, and stain nuclear with DAPI later. I am using the following conditions: ice-cold methanol fix for 5 min

However I found DAPI stain sometimes goes out of the nuclear and looks fluffy. I have no idea what is wrong could someone please help me,
thanks.

admin · #2 01-28-2009, 03:15 AM

Hi there, we often use formaldehyde or PFA for fixing cells (SHSY5Y). Why don't you try one of these ?
cheers

IoDiNE · #3 01-28-2009, 03:15 AM

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

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Methanol Fixation of SH-SY5Y cells for Immunofluorescence

Methanol Fixation of SH-SY5Y cells for Immunofluorescence - Immunofluorescence Forum

Methanol Fixation of SH-SY5Y cells for Immunofluorescence - Immunofluorescence Forum. Talk, discuss and post questions on immunofluorescence techniques including paraffin, frozen sections or cultured cells.