[Histonet] Using Isopropyl alcohol in tissue processor

[Rene J Buesa]

Iso-propanol (2-propanol) is the most secure, direct and cheapest alternative to xylene. The only thing I will recommend you is to have the first paraffin bath made of paraffin and 2-propanol at equal amounts in order to reduce the solubility gradient between the last 2-propanol and the first paraffin steps from 9 to 4.5 Mega Pascals.
Other than that you will not only eliminate xylene but will reduce costs since 2-propanol is cheaper than ethanol and xylene.
Most of the Peloris tissue processors users rely on 2-propanol as both dehydrating and "ante medium" agents.
To clean the tissue processor instead of xylene use a 2% aqueousus v/v solution of a laboratory strength dish washer soap, and you are set to have a tissue processing schedule free from xylene.René J.

--- On Sun, 1/4/09, Courtney Cain <Courtney.Cain <@t> pbrc.edu> wrote:


From: Courtney Cain <Courtney.Cain <@t> pbrc.edu>
Subject: [Histonet] Using Isopropyl alcohol in tissue processor
To: histonet <@t> lists.utsouthwestern.edu
Date: Sunday, January 4, 2009, 9:55 PM

 
I was wondering if anyone has tried Isopropyl Alcohol (2-propanol) as an
alternative to xylene substitutes in their tissue processors.  We are attempting
this method with the Thermo Excelsior to process mouse embryo, placenta, liver,
and mouse and rat organs. Currently our animal tissue has been immersion fixed,
but we have encountered extremely brittle and poor morphology of placenta and
livers. A few livers and kidneys have pulled away from the wax as well. 
 
Here is our basic protocol that has been attempted with and without vacuum.
 
Formaldehyde                  2 hr        
Formaldehyde                  2 hr
Isopropyl Alcohol 70%    20min
Isopropyl Alcohol 90%    20min
Isopropyl Alcohol 95%    20min
Isopropyl Alcohol100%  20min
Isopropyl Alcohol100%  20min
Isopropyl Alcohol100%  20min
Wax                                       30min
Wax                                       30min
Wax                                       30min  
 
Thanks,
 
Courtney Cain
Research Associate
Cell Biology & Bioimaging Core
Pennington Biomedical Research Center
225-763-2653
caincm <@t> pbrc.edu
 
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet