[Histonet] Alcian Blue Stabilizer & Manufacturing

Tue Apr 17 15:03:04 CDT 2007

Sarah, 
Perhaps, I was misunderstood.  I am speaking about
incorporating a post stabilizer to the lq.solution of
Alcian Blue, not the synthetic powder dye.  That is
what I am referring to for all the previous
correspondence.  Below was my protocol for making-up
Alcian Blue pH 2.5 w/o a post stabilizer.  

And by the way, alot has changed in Chuck's thoughts
of special stains since 1972.  I am BCC'ing him this
e-mail.

Believe it or not, in the beginning of my Biocare
association, I actually made all the IHC products,
H&E's, special stain solutions, QC'd them and bottled
them.  I learned alot about bulk manufacturing and
pH'ing. 

This is what I do with my Alcian Blue.  My "post"
stabilizer is missing from this protocol, because it
is proprietary.  By the way, Anatech has the best
Alcian Blue powder on the market! The best dye
content!

1.	Label the appropriately sized container with
description, lot number, storage conditions,
expiration date, prepared by and date.  Add a large
stir bar to the container.

2.	Measure the 3% acetic acid solution determined in
the Calculation section of Manufacturing Document. 
Transfer the 3% Acetic Acid to the labeled container,
and begin stirring on a large stir plate.  Add the
Alcian Blue and continue mixing solution.  

3.	Mix thoroughly for 30±5 minutes or until all the
chemicals and reagents have dissolved.  Visually
inspect solution to ensure adequate mixing.

4.	Measure the pH of the solution using a pH meter
calibrated at pH 1 and pH 4.  The pH must be 2.5 ±
0.1.  If necessary, adjust pH with NaOH of HCL.  

5.	The reagent is stable for 12 months at room
temperature.  Store the reagent at room temperature in
an opaque container.

6.	Test for quality control. 

> 
> Akemi
> 
--- Sarah Jones <Sarah.Jones <@t> dako.com> wrote:

> John A. Kiernan was my basic source for the research
> I did recently on
> Alcian Blue.  In his book, Histological &
> Histochemical Methods, he
> states (referring to Alcian Blue 8GS, 8GX or 8GN),
> "All these dyes are
> supplied mixed with boric acid, sodium sulphate, and
> dextrin." Also, in
> an e-mail to the Histonet, Kiernan writes:  "A low
> dye content such as
> the 10-20% in your old ICI material is not
> necessarily a bad thing for
> Alcian Blue, and batches with high dye content
> (70%+) often perform
> badly.  It seems likely that the additives (dextrin,
> boric acid, etc.)
> are needed in adequate amount."  In addition,
> quoting from Churukian et
> al, 1972: "Minimum dye content for Commission
> certification is 50%.  A
> high dye content is at the expense of less
> stabilizer.  Consequently,
> purer samples may have reduced shelf-lives."  
> 
> Sarah Jones
> 
> 
> 
> -----Original Message-----
> From: Akemi Allison-Tacha
> [mailto:akemiat3377 <@t> yahoo.com]
> Sent: Tuesday, April 17, 2007 11:49 AM
> To: Sarah Jones
> Subject: RE: [Histonet] Alcian Blue 8GX and S from
> someone who made it
> to sell
> 
> Very Interesting.  Chuck Churukian does not mention
> any stabilizers in
> his Special Stains book when making it up.  He was
> in charge of
> evaluating all dyes for the Biological Stains
> Commission before
> semi-retiring.
>  We spoke on several collaborations for Special
> Stains and he was one of
> the people who the CAT hematoxylin
> was named after.   
> 
> The image that is on the www.biocare.net site for
> Alcian Blue pH 2.5 was
> taken w/o the stabilizer.  I only started putting in
> a stabilizer after
> we started to sell it.
> 
> Akemi Allison-Tacha BS, HT (ASCP) HTL
> President
> Phoenix Lab Consulting & Staffing
> Specializing in Histology, SS, IHC, & Microarray
> Madison, WI
> Tele:           (925) 788-0900
> E-Mail: akemiat3377 <@t> yahoo.com
> 
> 
> --- Sarah Jones <Sarah.Jones <@t> dako.com> wrote:
> 
> > Unfortunately, with Alcian Blue a higher dye
> content doesn't
> > necessarily mean a better dye.  Alcian Blue
> absolutely NEEDS the
> > stabilizers in order to work properly.  The higher
> the dye content,
> > the fewer stabilizers are in the dye.  This is an
> exception to the
> > usual rule about dyes and high dye content.
> > Personally, I am finding that the synthetic Alcian
> Blue dye renders a
> > different blue than the Robin's egg blue we were
> used to seeing over
> > the years.  This blue seems to disappear if you
> use a Hematoxylin
> > counterstain, such as in the Alcian
> Blue/PAS-Hematoxylin.
> >
> > Sarah Jones
> > Dako North America
> >
> > -----Original Message-----
> > From: histonet-bounces <@t> lists.utsouthwestern.edu
> > [mailto:histonet-bounces <@t> lists.utsouthwestern.edu]
> > On Behalf Of Akemi
> > Allison-Tacha
> > Sent: Tuesday, April 17, 2007 11:19 AM
> > To: histonet <@t> lists.utsouthwestern.edu
> > Subject: [Histonet] Alcian Blue 8GX and S from
> someone who made it to
> > sell
> >
> > FYI to those of you who are not in the loop. 
> Alcian Blue has not been
> > procured in it's natural form for several years.
> > There were too many
> > fatalities during it's procurement from South
> America.  This came to
> > my attention in the late 90's.
> >
> > Anatech started creating it, in a synthetic form
> some years ago. It
> > was my understanding and I could be wrong, that
> for awhile, Anatech
> > was the only company who produced Alcian Blue to
> pass the Biological
> > Stains Commission evaluation process. It is my
> understanding, there is
> > only a 8GS being made now.  Again, I could be
> wrong.
> >
> > In the past few years, Dudley Chemical, Aldrich,
> Sigma, EM Science and
> > some Chinese companies were able to produce
> synthetic Alcian Blue,
> > although the dye content varies with each
> companies submission. You
> > should always get a seal from the Biological
> Stains Commission on each
> > bottle of Alcian Blue powder.  If the label is not
> on it, it has not
> > passed their evaluation process. This only means
> that it met it's
> > minimum standard.  The Commision will not divulge
> actual test results.
> >
> > When I started my R&D on Alcian Blue for Biocare,
> Anatech had the
> > highest dye content from the few companies that
> sold it.  It is only
> > my opinion, but they still have the best on the
> market.
> >
> > Akemi Allison-Tacha BS, HT (ASCP) HTL
> > President
> > Phoenix Lab Consulting & Staffing
> > Specializing in Histology, SS, IHC, & Microarray
> Madison, WI
> > Tele:         (925) 788-0900
> > E-Mail:       akemiat3377 <@t> yahoo.com
> >
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> 
> 