[Histonet] Acid Phosphatase Reaction

Rene J Buesa rjbuesa <@t> yahoo.com
Tue Oct 2 08:47:53 CDT 2012

Acid phosphatase is a standard procedure in FFPE (Formalin Fixed Paraffin Embedded) tissues. You will need to process your tissues accordingly.
Xylene is also standard procedure even when it should be avoided because of toxicity. There are many other alternatives to xylene usage.
I advise you to check in some TEM text that can answer your doubts.
René J.

From: "Duraine, Lita" <duraine <@t> bcm.edu>
To: "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu> 
Sent: Monday, October 1, 2012 6:35 PM
Subject: [Histonet] Acid Phosphatase Reaction 

Hello to all Histologists,
Perhaps someone can help me.  At our institution we primarily use electron microscopy for slides with toluidine blue, and grids for TEM.  The students here also do some immunostaining on their tissues.  We have a student though who would like to try using the Acid  Phosphatase Reaction to identify lysosomes as another alternative data to add to his paper.  Not having done any type of paraffin sectioning before how would we proceed?  I understand the basic tissue processing recipe with formalin, however does the procedure call for a special machine?  I noticed that all the recipes I have looked at online refer to using cassettes.  We primarily use microwave and vacuum to process our tissue in Karnovsky's for TEM.  Would the xylene cause any problems for us?  Are there any other stains that could work on resin embedded sections besides toluidine blue that would specifically stain lysosomes?  You advice would be greatly appreciated.

Thank you,

Lita Duraine
EM Technologist
Bellen Lab
HHMI- Molecular Genetics
Duncan Neurological Research Institute
1250 Moursund St.
Houston, TX 77030
Rm: N1165.17
MS: N1125.50
832-824-8772 TEM Room
979-549-6526 Cell

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