Re: mouse ovaries

From:Gayle Callis

Shorten your processing schedule - overdehydration, over clearing in xylene
(we now use Clearite 3 for all tissues, including decalcified bone) and
overexposure to heat of paraffin will cause dry, friable tissues.  It also
helps to face block, soak on ice block with water. You did not say what
kind of processor you have, but adding heat to processing through solvents
also is drying to murine tissue. 

You may want to try 
 30 min in 80% ethanol (skip the 70% ethanol(?), you have already done that
 2 changes 95% ethanol at 30 min per change
 2 changes 100% ethanol at 30 min per change
 2 changes Clearite 3 at 30 - 45 min per change (a good alternative is 1
change xylene, then 1 change      	Clearite 3 - this seems to help on
crunchieness issue)
 2 changes paraffin (try to use lower melting point i.e. 56C) 30 minutes
each change, a third change can be use. 

If you have a VIP or other automatic processor, program it so the total
time in 2 changes of 95% is spread out over three changes  (i.e. 15, 15, 30

If the tissue was fixed in 70%, it may be crunchier to begin with,
hopefully NBF was used prior to storage in 70%.  

30 At 10:28 AM 6/3/2003 -0500, you wrote:
>        A Histonet search revealed that you have been working with mouse
>tissue for a long time.  We have done mouse ovaries before but they have
>turned out crunchy.  I was wondering what you would suggest for a
>processing schedule.  The tissue will be in 70% alcohol.  We do not know
>any more details on fixation of the tissue.       Thank you for your time. 
>Margaret Perry  Lab Tech Histopath  South Dakota State University 
>Brookings SD 57007  605-688-5638          
Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
S. 19th and Lincoln St
Bozeman MT 59717-3610

406 994-6367 (lab with voice mail)
406 994-4303 (FAX)


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