[Histonet] frozen sections falling off slides

[Gayle Callis]

Stacey,

What I am not seeing here is if your tissues are prefixed with formalin or 
similar fixative before you do the cryosections, or are you working with 
unfixed tissue frozen sections?

Gayle Callis HTL, HT, MT(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717


You wrote:
   I'm cutting frozen rat scg at 10um in OCT. I cannot keep the sections 
from falling off the slides during the overnight incubation in primary 
anitbody. I have tried the Superfrost plus slides and currently we are 
using the Superfrost Plus GOLD slides which are supposed to be even better. 
THis is what I've tried so far:
   Slides on warmer for 1hr
   Slides on warmer for 2hr
   Slides on warmer for 5hr
   Slides on warmer for 2hr, then at RT overnight

   Sometimes ALL sections fall off. Other times just some of the sections 
will fall off.

   When I remove them from the warmer, I cover them with PBS to rehydrate. 
THen incubate overnight in my primary antibody at 4deg C.

   The next morning is when I notice all of the floaters. THey are usually 
foggy when the come out of the fridge. When I put the slides into PBS most 
if not all of the sections are gone.

   Please help! This is precious tissue and I can't loose anymore. I've 
never had this problem with any other tissue I've used. I dont believe its 
been a problem in the past in this lab either but noone knows what the 
difference is between then and now.

   Also, what setting do you use on the slide warmer (temp?) I've tried 
setting it between 2 and 3 (usually 40deg C)




Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)