[Histonet] H&E stain protocols

Dr. Michael Gudo (Morphisto GmbH) michael.gudo at morphisto.de
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

> 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
> The information contained in this e-mail message may be privileged, confidential, and/or protected from disclosure. This e-mail message may contain protected health information (PHI); dissemination of PHI should comply with applicable federal and state laws. If you are not the intended recipient, or an authorized representative of the intended recipient, any further review, disclosure, use, dissemination, distribution, or copying of this message or any attachment (or the information contained therein) is strictly prohibited. If you think that you have received this e-mail message in error, please notify the sender by return e-mail and delete all references to it and its contents from your systems.
> _______________________________________________
> Histonet mailing list
> Histonet at lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet

MORPHISTO Evolutionsforschung und Anwendung GmbH
PD Dr. phil. nat. Michael Gudo
Weismüllerstr. 45
60314 Frankfurt am Main
Telefon: 			069 / 400 3019 - 62
Telefax:			069 / 400 3019 - 64

E-Mail: michael.gudo at morphisto.de <mailto:info at morphisto.de>
Internet: http://www.morphisto.de/ <http://www.morphisto.de/>

Vertretungsberechtigter Geschäftsführer: Dr. Michael Gudo

Registergericht: Amtsgericht Frankfurt
Registernummer: HRB 74954
Umsatzsteuer-Identifikationsnummer gemäß § 27 a Umsatzsteuergesetz: DE243397199
Diese Nachricht ist ausschliesslich fuer den bezeichneten Adressaten oder dessen Vertreter bestimmt. Beachten Sie bitte, dass jede Form der unautorisierten Nutzung, Veroeffentlichung, Vervielfaeltigung oder Weitergabe des Inhaltes der Email nicht gestattet ist. Sollten Sie nicht der vorgesehene Adressat dieser Email oder dessen Vertreter sein, so bitten wir Sie, sich mit dem Absender der Email in Verbindung zu setzen und anschliessend diese Email und saemtliche Anhaenge zu loeschen.
This message is exclusively for the person addressed or their representative. Any form of the unauthorized use, publication, reproduction, copying or disclosure of the content of this e-mail is not permitted. If you are not the intended recipient of this message and its contents, please notify this sender immediately and delete this message and all its attachments subsequently.

More information about the Histonet mailing list