[Histonet] IHC with H & E staining

Elizabeth Chlipala liz at premierlab.com
Wed Jan 6 11:20:30 CST 2016


You can stain with H&E after DAB if you think it would help your how you analyze your slides.  DAB with the addition of a special stain has been published and for us depending upon the project that we are working on we may choose to use a different counterstain after an IHC that has DAB as an chromogen.  For example we have worked with macrophage markers counterstained with Prussian blue, another example is brain IHC markers counterstained Cresyl ect Violet rather than hematoxylin.  DAB is extremely stable and therefore many special stains or different counterstains can be used.  

If you are running an image analysis algorithm most canned algorithms are set up to function with a hematoxylin counterstain - you would need evaluate if a different counterstain would decrease the accuracy of your algorithm but you may have the ability to design an algorithm with the counterstain of your choice.


Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303) 682-3949 office
(303) 881-0763 cell
(303) 682-9060 fax
liz at premierlab.com

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From: Marcus Green via Histonet [histonet at lists.utsouthwestern.edu]
Sent: Wednesday, January 06, 2016 4:26 AM
To: histonet at lists.utsouthwestern.edu
Subject: [Histonet] IHC with H & E staining

Dear Users,

Happy New year - I hope this finds everybody well!

I was asked a very simple question yesterday - why don't you do H&E counterstaining on DAB stained samples. The question came about as we're looking at CD31 staining for blood-vessels and some look ruptured (we're keen to see red blood cells).

Instead of doing a consecutive cuts, a CD31 stain (Haem counterstain) on one slide and H&E on another, the question was asked why not do the Eosin stain as part of the counterstain....

I've never seen it done in the literature or in the clinic, and I've never asked why it's not done. Any assistance or advice would be greatly welcome - and my sincerest apologies if this is a very basic question?

Thanks in advance for your time,

kind regards,


Department of Oncology - University of Oxford,
Old Road Campus Research Building,
Roosevelt Drive,

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