[Histonet] H&E protocol question

Tony Reilly Tony_Reilly <@t> health.qld.gov.au
Mon Jul 25 19:27:31 CDT 2011


Hi Michelle
 
I have used both and my experience either is suitable for a good H&E if they are used appropriately.  For regressive staining you just need to optimise the time of haematoxylin staining with the concentration and time of the differentiation step.
 
In a previous position I used Mayer's which is a progressive stain.  We stained for 4 minutes in our H&E protocol however to get the best staining we gave it a "whiff" of differentiation (1 min with 0.025% Acid alcohol)  This was not to differentiate the nuclear staining but we found to get optimal nuclear staining the time required would result in some tissue types adopting a very pale blue haze in the background and removing this provided much better differential eosin staining.
 
For counterstaining special stains and IHC the Mayer's was by far the best I have used. 1 minute would provide delicate nuclear staining with a clear background.  This stain would not interfere with any other staining including nuclear staining for ER, PR, TTF1 etc.  Good bluing was achieved simply with a good wash in our quite basic tap water.
 
So I would recommend either for H&E but progressive for a counterstain.
 
regards
Tony
 
 
 
 

Tony Reilly  B.App.Sc. , M.Sc.
Chief Scientist, Anatomical Pathology
Pathology Queensland-PA Laboratory
_________________________________________________
Clinical and Statewide Services Division| QueenslandHealth
 
Level 1, Building 15,Princess Alexandra Hospital
Ipswich Road,WOOLLOONGABBA  Qld4102
Ph: 07 3176 2412
Mob: 0402 139411
Fax: 07 3176 2930
Email: tony_reilly <@t> health.qld.gov.au
Web:  www.health.qld.gov.au/qhcss/
 
 


>>> "histotech <@t> imagesbyhopper.com" <histotech <@t> imagesbyhopper.com> 7/25/2011 9:29 pm >>>
Hi Histonetters!

I'm looking for thoughts on preferences/pros/cons between using a progressive and a regressive H&E on routine daily work.

Which hematoxylins do you prefer (commercially prepared), which eosin?

Anyone have a tried and true protocol for each method?

Thanks!

Michelle

Sent from my iPhone
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