[Histonet] Mucin containing cells too blue

Wed Jan 23 15:09:58 CST 2008

Being the person who manufactured (CAT Hematoxylin)
for Biocare, Extremely Large Volumes by the way, I
would calibrate my pH meter daily using a #2 and #4
buffers.  You also must make sure that your probe is
designed for such a LOW pH.

During my learning curve, I also found that any
fluctuation in temperature, movement and also having
the pH meter close to any magnetic fields, could alter
the pH.  What I mean is, if your procedure requires
heat, the pH is quite different if it is slightly
warm, rather than if it is at RT.  I also found out
that if my pH meter was located to closely between (2)
magnetic stirrers. the pH would fluctuate.

When manufacturing, I would keep the pH probe in the
solution while it was being mixed.  After putting a
certain volume of GLACIAL ACETIC ACID in and mixing
it, I would let the solution rest, then recheck the
pH.  I would repeat this procedure till I reached my
desired pH.  The CAT hematoxylin also has glycerol in
the formula.  This was added after pH'ing.  The
glycerol raises the pH slightly.  There is also a
proprietary stabilizer incorporated.

Hope this helps and gives some insight.
Akemi Allison-Tacha

--- Tony Henwood <AnthonyH <@t> chw.edu.au> wrote:

> Good,
> I assume you would check the pH before you put it on
> the market.
> 
> Regards
> 
> Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys,
> CT(ASC)
> Laboratory Manager & Senior Scientist
> The Children's Hospital at Westmead,
> Locked Bag 4001, Westmead, 2145, AUSTRALIA.
> Tel: 612 9845 3306
> Fax: 612 9845 3318
> 
> 
> 
> 
> -----Original Message-----
> From: Akemi Allison-Tacha
> [mailto:akemiat3377 <@t> yahoo.com] 
> Sent: Wednesday, 23 January 2008 8:25 AM
> To: Tony Henwood; Greg Dobbin;
> Histonet <@t> lists.utsouthwestern.edu
> Subject: RE: [Histonet] Mucin containing cells too
> blue
> 
> 
> The pH of the Hematoxylin I developed for Biocare
> (CAT
> Hematoxylin) a progressive hematoxylin, was between
> pH
> 2.26 to 2.3.  I would let it oxidize (ripened) for 3
> months before releasing it for sale.  It would not
> over-stain mucin, especially in the GI tract.  
> 
> 
> --- Tony Henwood <AnthonyH <@t> chw.edu.au> wrote:
> 
> > Greg,
> > 
> > Don't assume that "fresh hematoxylin (and
> thererfore
> > correct pH) would
> > rectify the problem". Incorrect pH of the
> > Haematoxylin is the usual
> > problem. A feature of a haematoxylin solution that
> > has a high pH is the
> > staining of mucin, especially in the alimentary
> > tract. Decreasing the pH
> > of the staining solution to about 2.5 will
> decrease
> > this mucin staining.
> > 
> > Regards
> > 
> > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys,
> > CT(ASC)
> > Laboratory Manager & Senior Scientist
> > The Children's Hospital at Westmead,
> > Locked Bag 4001, Westmead, 2145, AUSTRALIA.
> > Tel: 612 9845 3306
> > Fax: 612 9845 3318
> > 
> > 
> > 
> > 
> > -----Original Message-----
> > From: histonet-bounces <@t> lists.utsouthwestern.edu
> > [mailto:histonet-bounces <@t> lists.utsouthwestern.edu]
> > On Behalf Of Greg
> > Dobbin
> > Sent: Wednesday, 23 January 2008 7:24 AM
> > To: Histonet <@t> lists.utsouthwestern.edu
> > Subject: [Histonet] Mucin containing cells too
> blue
> > 
> > 
> > Hi Folks,
> > I have been using the Surgipath Selectech reagents
> > (H&E) for almost 3
> > months now and I have not had a single complaint
> > about staining...until
> > now.  Suddenly the mucin containing cells are too
> > blue.  My first
> > thought was to make fresh Define (the Surgipath
> differentiator).  This
> > did not fix the problem.  Next I replaced the
> > hematoxylin with fresh
> > (not supposed to have to change before 2500
> slides,
> > but changed at 1500
> > slides to see if that would correct the problem).
> It
> > has not.  
> > 
> > I was sure that fresh hematoxylin (and thererfore
> > correct pH) would
> > rectify the problem.  What are the other usual
> > suspects here?  I should
> > also mention our H&E's are done on the Leica
> > Autostainer. I look forward
> > to receiving your collective insight! Greg
> > 
> > 
> > 
> > Greg Dobbin, R.T.
> > Chief Technologist, Histology Lab
> > Dept. of Laboratory Medicine,
> > Queen Elizabeth Hospital,
> > P.O. Box 6600
> > Charlottetown, PE    C1A 8T5
> > Phone: (902) 894-2337
> > Fax: (902) 894-2385
> > 
> > There is some merit in doing the right thing
> rather
> > badly,
> > but absolutely none in doing the wrong thing
> > excellently!
> > 
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> 
> Akemi Allison-Tacha, BS, HT(ASCP)HTL
> 
=== message truncated ===

Akemi Allison-Tacha, BS, HT(ASCP)HTL
President
Phoenix Lab Consulting & Staffing
Tele: (925)788-0900
E-Mail: akemiat3377 <@t> yahoo.com