[Histonet] H&E stain problems
Barry R Rittman
Barry.R.Rittman <@t> uth.tmc.edu
Wed May 19 09:30:07 CDT 2004
I personally do not like either Harris or Gill's hematoxylin as I was
brought up using hand staining and Ehrlich's hematoxylin made from
scratch. There are a lot of individuals using both these solution in
Histoland, so I have to assume that they can't all be wrong. There have
been a lot of hematoxylins formulated by individuals from several
cultures (not just dead white guys), after all hematoxylin was used in
South America for dyeing cloth way before anyone thought of using it for
staining tissue sections.
Recently Placido Domingo, who I think we can all agree is a great tenor,
when being interviewed on TV said "I do not understand rap but that is
my problem, I do respect it". I think that this is a valuable lesson
from a great master.
Let us not knock the younger generation just because they do not
necessarily have our principles or standards. It is arrogant of us to
regard ours as being the only ones possible or desirable. I do believe
that it is common sense to foster innovation. I am 65 but I find that I
can always learn something from "the new generation", life does not
necessarily end after 30.
Barry
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Connie
McManus
Sent: Wednesday, May 19, 2004 8:49 AM
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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