[Histonet] H&E stain protocols

Fri Oct 20 03:13:53 CDT 2017

Dear Toysha,

eosin stainings are soluble in lower alkohols, therefore the traditional procedure is to go shortly into 80 percent, just to remove surplus eosin but not more and then go directly in 96 (or 95) and 100 % alkohol. In our lab we use 96 % ethanol and then 2x 100 pesent 2-propanol.

Filtering of Eosin is useful if you stain many slides and the pH-value will change during staining. Since there are many different eosins etsablished in the labs (water-based eosins in 0.2, 0.5, 1.0 and 2.0 %, or ethanol based eosins in 0.2, 0.5, 1.0 or 2.0 % and these also with or without additional acetic acid to have pH values between 4.0 or 6.0), the protocols differ much from each other.

In our lab we use 1.0 % Eosin, in water or ethanol, and we change the solution (500 ml) after staining ~ 500 slides, and we do not filter, however, we check the pH-value each morning, and if this value is depart from 4,0 or 6,0 (depending on the results we want) we adjust the pH value with acetic acid and we filter the solution.

We use the Eosin which the chemical part of our company produces for the european market. So if you are interested, just contact me personally, we are now also able to deliver our products overseas.

With best regards
Michael

> Am 19.10.2017 um 23:46 schrieb Mayer,Toysha N via Histonet <histonet at lists.utsouthwestern.edu>:
> 
> We have a few questions to ask:
> In your H&E protocols, what is your set up after eosin: 70, 95, 100, 100, xylene  x 3or 95, 100, 100, 100, xylene x 3, or another variation?
> What brand of hematoxylin do you use? Eosin?
> Do you filter daily or at all? Eosin?
> How often do you change the hematoxylin? Eosin?
> Do you start with manufacturer suggestions for filtering and changing? Protocols? 
> Carson says one thing, Bancroft something else, and the manufacturer different. 
> I was taught 80, eosin, 70, 95, 100, 100, xylene x 3.
> We filtered on Wednesday, and changed and filtered on Monday. 
> This was homemade dyes though.
> When we switched to Richard Allan we no longer filtered when we changed and then only when a sheen showed up, usually Monday's. The reason was Richard Allan does not require filtering.  
> 
> Thanks,
> Toysha
> 
> 
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