[Histonet] Mouse pancreas

[Gayle Callis]

>Is the Prosoft a gradient?  Nothing wrong with your waterbath, it is the
processing schedule.  Your little mouse tissue is processed in a schedule
for much bigger tissues, and is probably overexposed to dehydrants.
Included is a schedule next to yours with explanation of equivalent to our
schedule, cut back times in dehydrants, clearant and paraffins and if still
friable, dry and cracked, cut back even more. Be sure you trim block and
soak in water, you can try warm then cold, or warm then ice water if
needed.  What you are doing is removing too much of the bound water
attached to tissue proteins along with free water in tissue spaces.   
>>
> 70% Alcohol- 30 min.    	     equivalant is 70% 30 min
>30 min >Prosoft- 1 hour 		"    "      80% 30 min
>15 min >Prosoft- 1 hour  		"    "      95% 15 min
>15 min >Prosoft- 1 hour  		"    "      95% 15 min
>30 min >Prosoft- 1 hour  		"    "      95% 30 min
>30 min >Prosoft- 1 hour  		"    "     100% 30 min
>30 min >Prosoft- 1 hour 30 min. 	"    "     100% 30 min
>45 min or 1 hour >Clearite 3- 1 hour 15 min.
>45 min or 1 hour >Clearite 3- 1 hour 15 min.
>elminate >Clearite 3- 1 hour 15 min. or adjust Clearite changes to be 20,
20, and 45 min
>30 min >Paraffin- 1 hour
>30 min >Paraffin- 1 hour
>30 min >Paraffin- 1 hour
>30 min >Paraffin- 1 hour
>for a total of 2 hours in paraffin at no more than 60C. 
>>
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 (lab with voice mail)
406 994-4303 (FAX)