[Histonet] aniline oil/Holzer stain

Paula Keene Pierce paula at excaliburpathology.com
Fri Oct 21 18:12:32 CDT 2016

You could try adding aqueous NaOH slowly to what you have until you start getting tiny white droplets in the water. Then they will fall and form an oily layer below the water.
Decant the water or use a separator funnel to drain the aniline oil out the bottom.

 Paula Keene Pierce, BS, HTL(ASCP)HTPresidentExcalibur Pathology, Inc.5830 N Blue Lake DriveNorman, OK 73069PH 405-759-3953FAX 405-759-7513www.excaliburpathology.com

      From: Susan Bachus via Histonet <histonet at lists.utsouthwestern.edu>
 To: histonet at lists.utsouthwestern.edu 
 Sent: Friday, October 21, 2016 5:28 PM
 Subject: [Histonet] aniline oil/Holzer stain
I'm trying to locate aniline oil for the Holzer glial fiber stain.  I 
purchased what was listed as aniline oil from ENG, but the first hint that 
something was wrong was that it was incompatible with the chloroform also 
included in the differentiating solution.  Looking at the fine print on the 
bottle, I saw that it was actually "aniline oil, 2.5% aqueous", which I 
would have interpreted as only containing 2.5% water, except that it seemed 
to be completely aqueous (2.5% aniline?), inasmuch as it wouldn't mix with 
the chloroform.  I can't find any other source online.  I see that it is 
also called aminobenzine or phenylamine, but am also having trouble locating 
sources under those names.  I realize aniline oil is relatively toxic, but 
am using it under a fume hood.  I tried dipping slides separately in the 
(apparently aqueous) aniline and then chloroform, but still didn't get good 
differentiation.  Can anyone suggest a source or a substitute?  Many thanks 
for your help!  Susan 

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