[Histonet] H&E stain problems

[Petia P Stefanova]

Hi Connie,

I use Leica autostainer XL and they offer even longer / 20 sec./
differentiation with acid alcohol in their manual.

Have a good day!
Petia



----- Original Message ----- 
From: "Connie McManus" <convmcm <@t> cc.usu.edu>
To: "'Petia P Stefanova'" <pstefanova <@t> sten.sunnybrook.utoronto.ca>; "'Megan
Kear'" <Megan.Kear <@t> hunter.health.nsw.gov.au>;
<histonet <@t> lists.utsouthwestern.edu>
Sent: Tuesday, May 18, 2004 10:09 AM
Subject: RE: [Histonet] H&E stain problems


> We use almost the exact same protocol... we use Surgipath Harris, but we
> prepare eosin in-house.  One thing I am amazed at in this protocol is
> the length of time in the acid alcohol.  Do you use an autostainer?  We
> have a Leica.  The time is set to 1 second in the acid ETOH and
> sometimes the sections are almost too differentiated.   I can't imagine
> 6 seconds in the acid ETOH!!  Even when I do H&E manually, I dip the
> slides in and quickly put them in running water.  I must have a very
> strong solution, I guess. Hmmmm. Interesting.  In truth, I prefer my
> hand stained sections better than when they're stained automatically.
>
> Just wondering and blabbering (hey, it's Tuesday, what do you expect??)
>
> Connie McManus
> Utah Veterinary Diagnostics Laboratory
> Utah State University
> Logan, UT
> Phone:  435/797-1891
> fax: 435/797-2805
>
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Petia P
> Stefanova
> Sent: Monday, May 17, 2004 6:45 AM
> To: Megan Kear; histonet <@t> lists.utsouthwestern.edu
> Subject: Re: [Histonet] H&E stain problems
>
> Hi,
>
> I use Harris's hematoxylin which is also regressive and purchase my
> hematoxylin and eosin /alcohol-based/ from www.surgipath.com.
> I get very good H&E staining with this protocol.
>
> REAGENT                       TIME
> Xylene                                3 min.
> Xylene                                3 min.
> Abs. alc.                              2 min.
> Abs. alc.                              2 min
> 95% alc.                              2 min
> 80% alc.                              2 min
> Wash /tap water/                  30 sec.
> Hematoxylin                          8 min.
> Wash /tap water/                  2 min.
> Acid alcohol                          6 sec.
> Wash /tap water/                  2 min.
> Wash /tap water/                  5 min
> 80% alc.                              30 sec.
> Eosin                                    15 sec.
> 95% alc.                              10 sec.
> Abs. alc.                              30 sec.
> Abs. alc.                              30 sec.
> Xylene                                1 min.
> Xylene                                1 min.
> Xylene                                Exit
>
> Hope it helps!
> Petia
>
>
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