[Histonet] RE: Tissue loss

[Liebenberg, Ellen]

We had the same problem with tissue coming of the slide after heat
treatment. Bone is particularly prone to loosening.  For our specimens
the fix was as follows:

 1.  After sectioning, dry the slides at 37 degrees overnight, then melt
the slides at 60 degrees for 30 minutes in a slide rack so they are
vertical (the paraffin should drip down the slide), and cool prior to
deparaffinization. 
 

2.   After the second deparaffinization (we use Clearite), air dry the
slides about 15 minutes, or until all the sections are completely free
of Clearite.  They should look white.  Then go into the third
deparaffinzation for the normal time.  We do 10 min per change.  Believe
it or not, drying during the deparafinnization steps does not impair
immunostaining. 
 

These suggestions came from some now frequent Histonet contributors in
response to my desperate phone calls back in the pre-Histonet days.

 

Hope this helps.

 

Ellen Liebenberg
University of California Medical Center
San Francisco, Ca
415-502-0809