[Histonet] Luxol Fast Blue tissue damage

[Mathew DeGutes]

Hey all any help is appreciated.  I am doing some myelin staining with luxol fast blue and have
found far too often that I get regular large "holes" in my tissue after staining.  My basic
protocol is this:

-Cryosections on slides rinse in ddH20 then PBS then 70% ethanol
-Place in .1% Luxol Fast Blue in 95% Ethanol and .05% Acetic Acid over night at 55C
-Remove from LFB and wash in ddH20 then PBS
-Differentiate in .05% Lithium Carbonate (usually about 30 sec)
-Wash in 70% ethanol and then counterstain or dehydrate and coverslip

The actual staining looks pretty good, but I just get these Round white holes as if something
bubbled up there and destroyed the tissue.  Nothing is boiling so I am curious if anyone else has
run into this problem and what might ameliorate it.  Thanks again.