[Histonet] Mouse Skin Processing

[Igor Deyneko]

Dear Histonetters!
I have been havins some problems with processing mouse skin. The blacks are
difficult to section, then tissue starts flowing away from the paraffin and
the sections do not retain Hematoxylin stain. That suggest to me that the
processing and paraffin infiltration were poor.
Here's the processing protocol I used for pieces no larger than 1cm^2.
70 % Alcohol - 1.5 hr
70 % Alcohol - 1.5 hr
70 % Alcohol - 1.0 hr
80 % Alcohol - 1.0 hr
95 % Alcohol - 1.0 hr
 95 % Alcohol - 1.0 hr
100 % Alcohol - 1.5 hr
 100 % Alcohol - 1.5 hr( Due to machine only having 2 stations for that
alcohol)
Xylene - 1.5 hr
 Xylene - 1.5 hr(Due to machine having only 2 stations for Xylene)
Paraffin - 1.0 hr
 Paraffin - 1.0 hr
 Paraffin - 1.0 hr

Any suggestions would be greately appreciated.
Thank you.
Igor Deyneko
Infinity Pharmaceuticals
Cambridge, MA.