[Histonet] Re: Tolivia, Navarro and Tolivia

Geoff McAuliffe mcauliff <@t> umdnj.edu
Mon Jun 25 09:21:05 CDT 2012

I have sympathy for your dilemma!
While I have no experience with the procedure in question there have 
been many, many publications since the 1960's trying to address this issue.
Personally, I have never seen one I though was much good/worth the 
effort so I have just stayed with toluidine blue in borax.
You might look at this paper by Charlotte Pool:
/Pool, C.R. Hematoxylin-eosin staining of OsO4-fixed epon embedded 
tissue; prestaining oxidation by acidified H2O2. Stain Technol.  
44:75-79, 1969./


On 6/25/2012 9:55 AM, Shanon Pink wrote:
> Hi Again Gayle,
> I'm sorry I was not very clear; I have already downloaded and read the
> publication, and I am trying the procedure out today. I was wondering
> about using a couple of substitutions and thought I might ask if
> anyone has tried this procedure before.
> I am on a four-week research project trying to find a nice
> polychromatic stain to use for epon-embedded thick sections for
> researchers here at the EM lab at St. Jude Children's Research
> Hospital in Memphis. Normally they just use Toluidine Blue, but in
> some cases researchers would like to have better differentiation on
> their thick sections. Initially we were trying to find a stain that
> approximates H&E since that's a familiar stain, but I've only had
> mediocre results with the following stains: (1) Toluidine Blue/Sodium
> Borate, followed with Basic Fuchsin, and (2) a homemade Paragon stain.
> The staining procedure described in the aforementioned publication is
> performed at  room temperature and is relatively fast. But as you
> point out, it involves phenol which is not fun to work with.
> Thanks to you and anyone else who has any thoughts!
> --Shanon Pink
> On Mon, Jun 25, 2012 at 8:42 AM, gayle callis<gayle.callis <@t> bresnan.net>  wrote:
>> Shanon,
>> Thank you.    If you are at a university, you should be able to get this
>> publication on line if their library can do this for you.   I am not sure I
>> can access it via my connections but will try later.   There are other
>> methylene blue/basic fuchsin protocols that work very well on epoxy
>> sections.  Keep in mind that the carbol component of this stain is just
>> another name for phenol, very toxic and often a controlled substance in
>> laboratories (can't be passed around!).
>> If you want I will go into my EM file for the other MB/BF methods, they are
>> simple to use and make up.   Let me know if you would like the publication.
>> Gayle Callis
>> -----Original Message-----
>> From: Shanon Pink [mailto:shanonpink <@t> gmail.com]
>> Sent: Monday, June 25, 2012 6:40 AM
>> To: gayle callis; histonet <@t> lists.utsouthwestern.edu
>> Subject: Re: Tolivia, Navarro and Tolivia
>> My apologies Gayle, I am new at this...
>> > From Histochemistry (1994) 101:51-55, "Polychromatic staining of epoxy
>> semithin sections: a new and simple method," by Tolivia, Navarro and
>> Tolivia. I am wondering if anyone out there has tried this technique.
>> Thank you for your time in responding!
>> --Shanon
>> On Fri, Jun 22, 2012 at 3:47 PM, gayle callis<gayle.callis <@t> bresnan.net>
>> wrote:
>>> What  journal, year and issue for this publication?
>>> ********************
>>> You wrote:
>>> Has anyone out there tried the staining procedure described in the
>>> paper "Polychromatic staining of epoxy semithin sections: a new and
>>> simple method," from 1994, by Tolivia, Navarro, and Tolivia? I would
>>> like to try it out on some thick sections embedded in Epon, but I
>>> can't find any carbol methylene blue and wondered about possible
>> substitutions.
>>> Gayle Callis
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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
mcauliff <@t> umdnj.edu

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