Protocol Online logo
Top : Forum Archives: : Cell Biology

cell fixation - How to fix cells? (Oct/15/2004 )

hi,everyone

I'd like to observe the cells using the Laser Scanning Confocal

Microscopy (LSCM), but I do not know how to dispose my cells before the

observation, that is to say, I want to know how to fix my cells effectively

so that the detectedcells could be observed much more truely.

Would you mind sharing with me if you know about it happenly?

Thanks a lot!

-geness-

Hi
What cells are you using?
we fix bacteria in 4% formaldehyde for 1 to 24 hours, works just fine for LSCM

-leahf-

hi,leahf

The cells used are HEK 293T cell lines.

-geness-

hi

u can use PFA 3% + gluta 0.5% 10 min at 37°C
rince with PBS or Tris 3*10min
u can store them 1 month at 4°C.

-sebwood-

Hi Geness

4 % paraformaldehyde in PBS will good for fixing cells.

Anil

-anilkumarpr-

hi,

Thanks a lot!

I believed that 4 % paraformaldehyde in PBS is good for fixing mammalian

cells.

But I'd like to know where I could get the manufacturer's instruction

about detailed fixing cells for Confocal detection.

If you know,would you mind sharing with me ? Thank you very much!

-geness-