[Histonet] Mucin containing cells too blue

[Akemi Allison-Tacha]

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
President
Phoenix Lab Consulting & Staffing
Tele: (925)788-0900
E-Mail: akemiat3377 <@t> yahoo.com