[Histonet] H&E stain problems
P. Emry
emry <@t> u.washington.edu
Wed May 19 12:36:29 CDT 2004
Ok, I have this pain that starts behind my right eye.
Let me get this straight...
I don't have to do 5 mins in clearing x3 100etoh x3 95etohx3
just 20-30 vigorus dips? Am I getting this?
then 20mins in whosoever's-hemotox,
15 mins in running water.
20-30 vigorus dips in 70etoh, 95x3,100ethox3,clearingx3 then slipcover.
Where does the acid alcohol come in and for howlong?
How did I miss this after all these years.
Thanks, and I knew Frank Sinatra....just thought I'd throw that in for the
music lovers here.
Trisha
Seattle
On Wed, 19 May 2004, Marshall Terry Dr, Consultant Histopathologist wrote:
> Close relative:-)
>
> Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path
> Consultant Pathologist
> Rotherham General Hospital
> South Yorkshire
> England
> terry.marshall <@t> rothgen.nhs.uk
>
> -----Original Message-----
> From: Connie McManus [mailto:convmcm <@t> cc.usu.edu]
> Sent: 19 May 2004 14:49
> To: 'Gary Gill'; Marshall Terry Dr, Consultant Histopathologist; 'Petia
> P Stefanova'; 'Megan Kear'; histonet <@t> lists.utsouthwestern.edu
> Cc: histonet <@t> lists.utsouthwestern.edu
> Subject: RE: [Histonet] H&E stain problems
>
>
> Gary
>
> Is Gill a relative or yours??? I don't give a hoot who made what when.
> I like Harris. I don't believe Gill has made any significant
> improvements over the old dead white guy's hematoxylin.
>
> BTW, classical music may have been composed by dead white guys, but I
> don't hear ANYTHING being composed today that has the depth, complexity
> and color of those great composers. I've studied piano from since I was
> 4 yrs old, and I've studied the organ (I've played as a church
> organist)for almost 20 years. I know music like the back of my hand
> and I love ALL music (except gangstah stuff). SO DON'T try to tell me
> about what music is living and worth my while.
>
> Dead white Europeans. Yeah, right. I'd like to see someone between 8 and
> 14 in this generation compose music like Felix Mendelsohnn's or
> Mozart's.
>
> Connie McManus
> Utah Veterinary Diagnostics Laboratory
> Utah State University
> Logan, UT
> Phone: 435/797-1891
> fax: 435/797-2805
>
>
> -----Original Message-----
> From: Gary Gill [mailto:garygill <@t> dcla.com]
> Sent: Tuesday, May 18, 2004 10:04 AM
> To: 'Connie McManus'; 'Marshall Terry Dr,Consultant Histopathologist';
> 'Petia P Stefanova'; 'Megan Kear'; histonet <@t> lists.utsouthwestern.edu
> Cc: histonet <@t> lists.utsouthwestern.edu
> Subject: RE: [Histonet] H&E stain problems
>
> You may have heard that classical music was composed by dead white
> Europeans. Well, Harris hematoxylin was composed by a dead white
> American
> (physician at Jefferson Hospital in 1904). Gill hematoxylin was
> composed by
> a live white American. So if you want to liven things up, go with
> Gill's!
>
> Gary Gill (one and the same)
>
> PS -- No royalties involved, thanks to bad advice in 1972 from corporate
> counsel for Johns Hopkins Medical School.
>
> -----Original Message-----
> From: Connie McManus [mailto:convmcm <@t> cc.usu.edu]
> Sent: Tuesday, May 18, 2004 10:41 AM
> To: 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P
> Stefanova';
> 'Megan Kear'; histonet <@t> lists.utsouthwestern.edu
> Cc: histonet <@t> lists.utsouthwestern.edu
> Subject: RE: [Histonet] H&E stain problems
>
>
> Wow. What a lot of interesting comments!!
>
> I agree with Terry re the agitation. When I watch the stainer do those
> dips
> (I can program how many, but NOT the briskness), I wonder if you could
> even
> call it agitation. My hand dips are very brisk. Also, I don't bother
> letting the slides stay in the alcohols for 1 minute or 2, I give the
> slides
> about 20 -30 good brisk dips in each solution, then the timed rinses &
> staining. This has always been far more satisfactory to me than those
> sllllooooowwwww dips from the machine.
>
> As for the kind of hematoxylin, someone suggested I throw out the Harris
> and
> do Gills III. I've tried Gill III before and I much prefer the Harris.
> So
> it's just a matter of personal preference on that... AND what your
> pathologist likes *G*
>
> Everyone having an nice Tuesday??? *g*
>
> Connie McManus
> Utah Veterinary Diagnostics Laboratory
> Utah State University
> Logan, UT
> Phone: 435/797-1891
> fax: 435/797-2805
>
>
> -----Original Message-----
> From: Marshall Terry Dr, Consultant Histopathologist
> [mailto:Terry.Marshall <@t> rothgen.nhs.uk]
> Sent: Tuesday, May 18, 2004 7:35 AM
> To: Connie McManus; Petia P Stefanova; Megan Kear;
> histonet <@t> lists.utsouthwestern.edu
> Subject: RE: [Histonet] H&E stain problems
>
> Connie remarks:
>
> "In truth, I prefer my hand stained sections better than when they're
> stained automatically."
>
> When I first saw what I call x-y stainers, I thought that we had in
> this,
> something that reproduced hand staining. Not so. What I think is lacking
> is
> the brisk agitation necessary to break down the interface between old
> and
> new solution. I'm not so sure about the rinsing either.
>
> Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant
> Pathologist Rotherham General Hospital South Yorkshire England
> terry.marshall <@t> rothgen.nhs.uk
>
> -----Original Message-----
> From: Connie McManus [mailto:convmcm <@t> cc.usu.edu]
> Sent: 18 May 2004 15:09
> To: 'Petia P Stefanova'; 'Megan Kear'; histonet <@t> lists.utsouthwestern.edu
> 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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