3 replies to this topic

[Goplanka]

Hi,
can anyone advice me how to prepare and fix non-adherent cells on slides to the peroxidase immunostaining? I wonder, if I dry them on slides and than fix in paraformaldehyde, won't them give too high background? Thanks for help.
Margaret

[wirly]

The only way I've doen this is with a Cyto-Spin which just centrifuges the cells down onto a slide while removing most of the liquid and you can then fix then or whatever you want.

[biomed]

Goplanka, on Sep 30 2004, 03:24 AM, said:

Hi,
can anyone advice me how to prepare and fix non-adherent cells on slides to the peroxidase immunostaining? I wonder, if I dry them on slides and than fix in paraformaldehyde, won't them give too high background? Thanks for help.
Margaret

Hi,
I used to fix cell first with 4% ice-colded paraformaldehyde for 15 min, then smear the cells on slide and let it dry.
Good lock.
Biomed

[littlecell]

i used to stain the sperm (they are able to swim), which are non-adherent. i think you can fix them at first in 4% PFA, then block, add primary antibody, wash, incubate them with seconday antibody, wash, suspend them, all these steps could be done in a 1.5ml tube (save you antibody). finally  pipette one drop (about 30ul cell suspension)  on the slide, put on a microscope cover glass, press  and observe under the immunofluorescence microscope. it worked very well on sperm.  good luck!