[Histonet] Help in brain tisuue processing

[Leila Ramos]

Dear Mr. or Ms
We are working in a small lab from Canarias Island (Spain). We work with
goat brain with manual procedure, our problems are several in the
processing:

   1. We obtain very dry blocks, when we try to cut with the microtome,
    the tissues are fragile and it is impossible to obtain the correct tissue.
   2. When "we get proper block" (we can cut better the tissue), then we
   try to put in the float, the tissue appears like a white colour in the
   slide. After when we finish the stain process (hematoxyline-eosina and
   kluver barrera both of them) the tissues are not attached on the glass
   slides.

I explain you in fast way our protocol:

   1. Fixation: the whole brain is embedded in Formol 10% (1 week
   minimum)
   2. Dehydration: 80º etanol 30 minutes; 96º 30 min x 2 times; 100 º 30
   min x 5 times; xylene 1 hour x 2 times
   3. Tissue embedding in paraffin. In cassetes with pure paraffin all
   night- Only 1 step other times 2 changes in paraffin 1 hour
   4. Cut with microtome at 8 um, slide with polylisine.
   5. Stain process (Hematoxyline-Eosine and kluver barrera)

Thank a lot!!

Leila Ramos
Instituto de Investigación y Ciencias de Puerto del Rosario