[Histonet] Need technique information

[Johnson, Teri]

Hi all,

I have some slides of cultured cells which are immunostained, fixed in
4% PFA and sorted through a FACS. The sorted cells are then dropped onto
glass slides for H&E and eventually IHC and ISH staining. They have
dropped some cell suspension on to a plus-slide and air dried at 60
degrees C, then post-fixed in formalin and air dried again. The
morphology isn't very good and the cytoplasm doesn't show up at all on
H&E. Does anybody have a protocol for doing either cytospins (may not be
possible, only 2000 cells total!) or cell suspension dried slides to get
better morphology?

I believe they probably don't need heat to dry them. I also believe they
don't need an additional post-fix, but if they do one, to keep the slide
wet afterward (no additional air-drying step) and begin whatever
staining technique follows.

Thanks in advance for any suggestions!

Teri Johnson, HT(ASCP)QIHC
Managing Director Histology Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, MO 64110