RE: Daily Digest
| From: | Donella.Stillings@UCHSC.edu |
To LC:
TIA-1 can be found from Santa Cruz Biotechnology, 1-800-457-3801.
Hope this helps.
Donella
-----Original Message-----
From: HistoNet Server
[mailto:histonet@pathology.swmed.edu]
Sent: Thursday, October 25, 2001 11:54 PM
To: HistoNet Server
Subject: Daily Digest
----------------------------------------------------------------------
Date: 25 Oct 2001 02:25:26 -0500
From: Mikael Niku
Subject: Re: fixation procedures for cytology immunocytochem
Our procedure for cytospins & smears intended for
IHC or/and ISH (in situ hybridization):
- - air dry (at least a few hours after preparing)
- - fix in 100% ethanol at -20C for 30 min OR
in acetone at -20C for 10 min
- - air dry (at least 30 min)
- - store at -80C in small airtight containers (5 slides
each)
We don't use nuclear antigens currently but as genomic ISH
works there seems to be no permeability problems.
We switched to storage at -80C after problems with samples
stored at -20C.
\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\
Mikael Niku URL: www.helsinki.fi/~mniku/
University of Helsinki Dept. Basic Veterinary Sciences
- Mitako mielta olen lansimaisesta sivistyksesta?
Minusta se olisi erinomainen ajatus!
- Gandhi
////////////////////////////////////////////////////////////
- ----- Original Message -----
From: "David Grehan"
To: "'histonet'"
Sent: Thursday, October 25, 2001 1:49 AM
Subject: fixation procedures for cytology immunocytochem
> Hi histocybernetters
> I would like to get your opinions on procedures for
cytology specimens
> that are prepared for immunocytochemistry. I am currently
fixing direct
> smears or cytospins immediately in 95% absolute alcohol
for 10 mins .I
> do this for any cytoplasmic or cell membrane antigens . I
fix in
> methanol:acetone 50:50 for 10 mins for any nuclear
antigens such as Tdt
> or Myo D1 (to increase permeabilisation)is this
necessary?. I work in a
> busy paediatric laboratory so we don't get a lot of
cytology specimens
> but on occasion we are requested to do them. My results to
date have
> been patchy especially following storage at -20C. I would
like to be a
> bit more confident about my procedures and would like any
comments from
> anyone doing a lot of immunocytochemistry on cytologyy
specimens . Also
> how do you store your smears ? Do you spray fix or use
PEG.?
>
>
>
----------------------------------------------------------------------
Date: 25 Oct 2001 03:09:12 -0500
From: Jenny Molde
Subject: Dapi
Hi,
We are looking for any information on how to make up the
dapi counterstain
for nuclei using Immunofluoresence? Any info will be greatly
appreciatted.
Many thanks.
Jenny Molde
University of Cape Town
Observatory
Cape Heart Centre
ctsmolde@samiot.uct.ac.za
----------------------------------------------------------------------
Date: 25 Oct 2001 04:51:44 -0500
From: NLOUISEA@aol.com
Subject: forensic pathologist
Good morning,
A friend of mine, an attorney, is looking for a forensic
pathologist in the
New England (he's in MA) area that does expect opinion legal
work. I thought
someone in this very resourceful group might know of
someone. Please reply
off-net so as not to clutter the space.
Thanks
Nancy
----------------------------------------------------------------------
Date: 25 Oct 2001 07:35:34 -0500
From: Barry Rittman
Subject: OT
A former student who is in the US navy came to visit us. He
told us of
an incident about three weeks ago when his ship was being
passed by a
British destroyer.
The sailors on the destroyer appeared on deck in full dress
uniforms,
saluted the US ship and ran up the Stars and Stripes.
While I was born England I have come to love and respect
America and
what both countries stand for.
I found this gesture very touching and although I cannot
thank the
sailors directly I would like to thank Britons as a whole
for their
support in these difficult times.
Thank you.
Barry
----------------------------------------------------------------------
Date: 25 Oct 2001 09:21:50 -0500
From: "Sebree Linda A."
Subject: RE: fixation procedures for cytology immunocytochem
Hi David,
Our cytology department routinely fixes any cytology
preparations destined
for IHC in acetone for 3.5 minutes. This seems to work for
most immuno
stains. Several exceptions are ER & PR; these seem to need
special
attention such as fixation in Zamboni's fixative or the
3-step 10%
NBF/methanol/acetone fixation however we still haven't found
a reproducible
fixation protocol. On at least one occasion we have achieved
positive
staining of acetone fixed cytospins for ER and PR but again
we have not been
able to reproduce this. One other exception is Calretinin;
although
fixation is the same acetone protocol, we do a heat
pretreatment of 2" in
the Biocare DeCloaking chamber with Biocare's Nuclear
DeCloaker buffer or
20" in a 93 degree C waterbath in the same buffer. Dana
Dittus suggested
heat pretreatment and explained to me the mechanism involved
(its not really
the same antigen retrieval mechanism one usually associates
with FFPE
tissues) but she would be better able to explain this than
I.
Our cytopathologists are more and more going with cell
blocks when there is
enough material to make one and this suits us just fine as
we know how to
treat these.
Linda A. Sebree, HT(ASCP)
University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
A4/204-2472
600 Highland Ave.
Madison, WI 53792-2472
(608)265-6596
FAX: (608)262-7174
- -----Original Message-----
From: David Grehan [mailto:david.grehan@olhsc.ie]
Sent: Wednesday, October 24, 2001 5:50 PM
To: 'histonet'
Subject: fixation procedures for cytology immunocytochem
Hi histocybernetters
I would like to get your opinions on procedures for cytology
specimens
that are prepared for immunocytochemistry. I am currently
fixing direct
smears or cytospins immediately in 95% absolute alcohol for
10 mins .I
do this for any cytoplasmic or cell membrane antigens . I
fix in
methanol:acetone 50:50 for 10 mins for any nuclear antigens
such as Tdt
or Myo D1 (to increase permeabilisation)is this necessary?.
I work in a
busy paediatric laboratory so we don't get a lot of cytology
specimens
but on occasion we are requested to do them. My results to
date have
been patchy especially following storage at -20C. I would
like to be a
bit more confident about my procedures and would like any
comments from
anyone doing a lot of immunocytochemistry on cytologyy
specimens . Also
how do you store your smears ? Do you spray fix or use PEG.?
----------------------------------------------------------------------
Date: 25 Oct 2001 09:22:10 -0500
From: "Weems, Joyce"
Subject: RE: OT
Amen - such a change from the Vietnam era - here and
everywhere!
Thanks from me too.
Joyce Weems
Pathology Manager
Saint Joseph's Hospital of Atlanta
-----Original Message-----
From: Barry Rittman
[SMTP:brittman@mail.db.uth.tmc.edu]
Sent: Thursday, October 25, 2001 8:00 AM
To: histology
Subject: OT
A former student who is in the US navy came to visit
us. He told us
of
an incident about three weeks ago when his ship was
being passed by
a
British destroyer.
The sailors on the destroyer appeared on deck in
full dress
uniforms,
saluted the US ship and ran up the Stars and
Stripes.
While I was born England I have come to love and
respect America and
what both countries stand for.
I found this gesture very touching and although I
cannot thank the
sailors directly I would like to thank Britons as a
whole for their
support in these difficult times.
Thank you.
Barry
----------------------------------------------------------------------
Date: 25 Oct 2001 09:34:22 -0500
From: Connie McManus
Subject: Re: OT
Barry,
Thank you for sharing this! I am a born and bred American,
lived in the
wild west most of my life, but I have a great love for the
Brits, too, and
am DEEPLY greatful for their support of our President in
these times...
THANKS TO THE UK!! *G*
Up with the Union Jack!
Connie McManus
At 06:59 AM 10/25/01 -0500, Barry Rittman wrote:
>A former student who is in the US navy came to visit us. He
told us of
>an incident about three weeks ago when his ship was being
passed by a
>British destroyer.
>The sailors on the destroyer appeared on deck in full dress
uniforms,
>saluted the US ship and ran up the Stars and Stripes.
>While I was born England I have come to love and respect
America and
>what both countries stand for.
>I found this gesture very touching and although I cannot
thank the
>sailors directly I would like to thank Britons as a whole
for their
>support in these difficult times.
>Thank you.
>Barry
>
>
>
Veterinary Diagnostics Lab
Utah State University
Logan, UT
USA
(435) 797-1891
fax (435) 797-2805
----------------------------------------------------------------------
Date: 25 Oct 2001 09:34:45 -0500
From: "Weems, Joyce"
Subject: FW: Personnel Shortage
This is an email I received from my Director regarding the
Congressional
Bill. Perhaps it will help clarify the email sent by Glenda
Hoye
.
The bill does specifically mention Histology and Cytology.
Send your response to ElissaP@ascls.org
or call or fax as below.
We all need to speak out! Next year is a BIG election year.
Use that to our
advantage. We need to speak directly to our Senators and
Representatives
also - emails, voice mails, etc.
I belong to the Advocacy Committee - I will forward things
along as I see
relevance. (Excuse us, Dear Friends outside the USA!)
God bless us everyone!
Joyce Weems
Pathology Manager
Saint Joseph's Hospital of Atlanta
Subject: Personnel Shortage
Hi Everyone,
As this Congress winds down its work, there is some concern
that H.R. 1948
will not be enacted. In addition, since there never was a
Senate bill, it
is unlikely that any of it would be implemented. In
meetings that we
(ASCLS) have been having with the College of American
Pathologists, we have
discussed what it would really take to fix the shortage.
The College's
contacts on the Hill have indicated that they believe that
there is a
shortage but that loan forgiveness programs aren't really
going to fix the
problem. David Fowler reported that Secretary Thompson
(HHS), as part of a
video conference on anthrax, stated four different times
about the
difficulty the labs were having with keeping up with the
amount of work and
stated that this was a concern that was brought to his
attention and needs
to be addressed. He also stated that he needs some direction
about the best
way to help the labs. As David said, this is the time for
us to tell folks
what is needed for the short and long term to make sure that
the public gets
the level of testing and quality that they deserve. CAP
agrees and is
trying to get a meeting with the Secretary for the two
organizations (and
other organizations that might want to attend). CAP is
asking us to come up
with some really effective strategies to fix this shortage
so that CAP can
endorse them and we can all be sending the same message.
Therefore, we need
to do some brainstorming. We know that salary and
recognition are two
crucial contributors to the shortage. But are there other
things? And what
is the answer?
So, please talk among yourselves, share this email with as
many folks as you
would like but get back to David and/or I with:
a. what is contributing to the shortage (with some
prioritization of the
list)
b. what will really fix it? (including money - but money
where - to just
the hospitals, to education programs, to ? where)
Please get back to us as soon as possible but no later than
Oct 31. Thanks!
Elissa Passiment
ASCLS Executive Vice President
301.657.2768, X3015
301.657.2909 (fax)
7910 Woodmont Ave, #530
Bethesda, MD 20814-3015
----------------------------------------------------------------------
Date: 25 Oct 2001 09:35:06 -0500
From: Linda Jenkins
Subject: H&E for plastic
Good Morning, Rachel!
You are quite observant on the comment concerning
the hematoxylin
precipitate you observed on the surface of the plastic
embedded slide --
this is precisely what happens with the routine paraffin
"H&E" when applied
to plastic sections. Gayle and Evan are both correct when
mentioning the
methylene blue/basic fuchsin stain as an excellent
substitute. This stain
requires etching. However, not all plastics etch the same.
Methyl
methacrylate sections can be etched with a weak hydrochloric
acid solution
whereas a Technovit 7200 embedded slide will crack
profusely. These slides
have to be etched with a weak formic acid solution.
So...several factors
to consider; use the methylene blue/basic fuchsin instead of
H&E (I will be
glad to send you my version as well), which plastic used
will influence the
etching solution, and, as Gayle said, do not coverslip with
mounting
media. I actually have found that adding a drop of
immersion oil and then
coverslippping before photography enhances the picture.
Remove coverslip
afterwards and remove oil with a Kimwipe.
Best wishes,
Linda
P.S. For Evan , et al.: Cracking of certain plastic
embedded sections is
also a problem with light curing monomers such as Technovit
7200. These
must be stored in the dark as room light will continue to
slowly polymerize
the section causing cracking and bowing of the plastic
section.
----------------------------------------------------------------------
Date: 25 Oct 2001 09:53:22 -0500
From: "Johnson, Theresa"
Subject: Re: antigen retrieval
Tim Morken wrote: >>I'm wondering, are they worried about
how formalin will
affect their
protiens? It seems that would be a bigger problem!<<
Yes, they are worried about how formalin will affect the
proteins. When
they suspect that to be a problem, we do IHC in frozen
section.
I read somewhere that there is some speculation that the
proteins are often
protected by the fixative, which is why we can unmask them
using AR. I guess
we've learned that heat isn't as much a factor in destroying
them as first
thought, though I'm sure some are adversely affected.
Teri Johnson
Manager Histology Core Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, Missouri 64110
tjj@stowers-institute.org
----------------------------------------------------------------------
Date: 25 Oct 2001 11:31:55 -0500
From: Bruce Gapinski
Subject: RE: Personnel Shortage
Does this mean another raise for Cytology? Our Cytology
department got a 30%
raise last year! We got 2%. I'm sick of it! <-- sorry,
venting.
Respectfully,
Bruce Gapinski HT(ASCP)
-----Original Message-----
From: Weems, Joyce
[mailto:JWEEMS@sjha.org]
Sent: Thursday, October 25, 2001 6:40 AM
To: 'Histonet'
Subject: FW: Personnel Shortage
This is an email I received from my Director
regarding the
Congressional
Bill. Perhaps it will help clarify the email
sent by Glenda
Hoye
.
The bill does specifically mention Histology
and Cytology.
Send your response to ElissaP@ascls.org
or call or fax as below.
We all need to speak out! Next year is a BIG
election year.
Use that to our
advantage. We need to speak directly to our
Senators and
Representatives
also - emails, voice mails, etc.
I belong to the Advocacy Committee - I will
forward things
along as I see
relevance. (Excuse us, Dear Friends outside
the USA!)
God bless us everyone!
Joyce Weems
Pathology Manager
Saint Joseph's Hospital of Atlanta
Subject: Personnel Shortage
Hi Everyone,
As this Congress winds down its work, there
is some concern
that H.R. 1948
will not be enacted. In addition, since
there never was a
Senate bill, it
is unlikely that any of it would be
implemented. In
meetings that we
(ASCLS) have been having with the College of
American
Pathologists, we have
discussed what it would really take to fix
the shortage.
The College's
contacts on the Hill have indicated that
they believe that
there is a
shortage but that loan forgiveness programs
aren't really
going to fix the
problem. David Fowler reported that
Secretary Thompson
(HHS), as part of a
video conference on anthrax, stated four
different times
about the
difficulty the labs were having with keeping
up with the
amount of work and
stated that this was a concern that was
brought to his
attention and needs
to be addressed. He also stated that he
needs some direction
about the best
way to help the labs. As David said, this
is the time for
us to tell folks
what is needed for the short and long term
to make sure that
the public gets
the level of testing and quality that they
deserve. CAP
agrees and is
trying to get a meeting with the Secretary
for the two
organizations (and
other organizations that might want to
attend). CAP is
asking us to come up
with some really effective strategies to fix
this shortage
so that CAP can
endorse them and we can all be sending the
same message.
Therefore, we need
to do some brainstorming. We know that
salary and
recognition are two
crucial contributors to the shortage. But
are there other
things? And what
is the answer?
So, please talk among yourselves, share this
email with as
many folks as you
would like but get back to David and/or I
with:
a. what is contributing to the shortage
(with some
prioritization of the
list)
b. what will really fix it? (including
money - but money
where - to just
the hospitals, to education programs, to ?
where)
Please get back to us as soon as possible
but no later than
Oct 31. Thanks!
Elissa Passiment
ASCLS Executive Vice President
301.657.2768, X3015
301.657.2909 (fax)
7910 Woodmont Ave, #530
Bethesda, MD 20814-3015
----------------------------------------------------------------------
Date: 25 Oct 2001 11:32:20 -0500
From: kpawlow@swbell.net
Subject: IHC of CK18 :(
Help,
I am having trouble getting any staining (background or
otherwise) with
CK18 labeling. The tissue is formalin fixed, paraffin
embedded rat sub-
mandibular gland, which according to the data sheet should
stain.
I am using a Sigma antibody at the recommended dilution and
twice that
strength, for the recommended time (1 hr) and at 4C
overnight. I have
tried protease E digestion and micro waving with citrate
phosphate
buffer to no avail. I have tried the MOM kit from Vector (I
wanted to
eventually do mice) and a direct labeled secondary with no
blocking.
All I've seen so far is RBC staining (endogenous) on the
tissue that
I didn't first expose to H2O2. I figure I should at least
see some
nonspecific staining of the secondary when I skip all
blocking steps,
but that doesn't happen. Any ideas? [I checked the DAB and
secondary
HRP and they seem to react with one another.]
Also, we used a xylene substitute to deparaffinize
(histoclear).
Anyone had problems with using xylene substitutes with
IHC? Am I missing some-thing obvious?
I've contacted both Sigma and Vector, but so far the only
thing they
suggest that I haven't already tried was xylene
deparaffinization.
Karen Pawlowski, Ph.D.
----------------------------------------------------------------------
Date: 25 Oct 2001 11:32:36 -0500
From: Andrea Grantham
Subject: ? for laser capture gurus
I have run into a problem with some frozen sections I did
for laser capture
microdissection and I'm wondering if anything can be done to
salvage the
slides that I cut (many slides!).
When I did the frozens I picked up the tissue on my usual
slides which are
the Snowcoat X-tra slides from Surgipath. Now that the
tissue is "nailed"
to the glass it won't come loose and allow itself to be
captured. Is there
anything that can be done or is it back to the cryostat?
Thanks!!
Andi Grantham
.....................................................................
: Andrea Grantham, HT(ASCP) Dept. of Cell Biology &
Anatomy :
: Sr. Research Specialist University of Arizona
:
: (office: AHSC 4212) P.O. Box 245044
:
: (voice: 520-626-4415) Tucson, AZ 85724-5044
USA :
: (FAX: 520-626-2097) (email:
algranth@u.arizona.edu) :
:...................................................................:
http://www.cba.arizona.edu/histology-lab.html
----------------------------------------------------------------------
Date: 25 Oct 2001 13:01:30 -0500
From: Abizar Lakdawalla
Subject: Re: IHC of CK18 :(
Hi Karen, if the antibody is clone DC-10, then antigen
retrieval might work
better than protease digestion. A buffer of pH 4 gave us the
best
signal-to-noise.
Abizar
www.innogenex.com
Karen Pawlowski wrote:
> Help,
>
> I am having trouble getting any staining (background or
otherwise) with
> CK18 labeling. The tissue is formalin fixed, paraffin
embedded rat sub-
> mandibular gland, which according to the data sheet should
stain.
> I am using a Sigma antibody at the recommended dilution
and twice that
> strength, for the recommended time (1 hr) and at 4C
overnight. I have
> tried protease E digestion and micro waving with citrate
phosphate
> buffer to no avail. I have tried the MOM kit from Vector
(I wanted to
> eventually do mice) and a direct labeled secondary with no
blocking.
>
> All I've seen so far is RBC staining (endogenous) on the
tissue that
> I didn't first expose to H2O2. I figure I should at least
see some
> nonspecific staining of the secondary when I skip all
blocking steps,
> but that doesn't happen. Any ideas? [I checked the DAB and
secondary
> HRP and they seem to react with one another.]
>
> Also, we used a xylene substitute to deparaffinize
(histoclear).
> Anyone had problems with using xylene substitutes with
> IHC? Am I missing some-thing obvious?
>
> I've contacted both Sigma and Vector, but so far the only
thing they
> suggest that I haven't already tried was xylene
deparaffinization.
>
> Karen Pawlowski, Ph.D.
----------------------------------------------------------------------
Date: 25 Oct 2001 13:01:54 -0500
From: Lynde77627@aol.com
Subject: MyoD1
I saw a note several weeks ago requesting help with MyoD1
but never saw any
responses. I am now in the process of attempting to get it
to work. If
anyone has any insite to this antibody I sure would
appreciate it. I am using
FFPE tissue and NovaCastra antibody.
Thanks in advance.
Linda
----------------------------------------------------------------------
Date: 25 Oct 2001 13:02:19 -0500
From: "L. Muffley"
Subject: IHC web site resources
For the person interested in general info about IHC,
here are some web resources to suppliment your knowledge:
http://www.chemicon.com/resource/a2D.htm
http://immuno.hypermart.net/
http://www.protocol-online.net/immuno/immunohisto/immunohistochemistry.htm
http://www.rndsystems.com/asp/g_sitebuilder.asp?bodyId=236
Lara Muffley
Dermatology Dept
University of Washington
Seattle, WA
muffley@u.washington.edu
----------------------------------------------------------------------
Date: 25 Oct 2001 13:15:10 -0500
From: Julie Collins
Subject: microtome help
Can anyone tell me where I can get a decent inexpensive, new
microtome? I
haven't purchased one in 10 years and have no idea where to
start looking. I
don't need anything fancy. My favorite microtome I had was
the old black AO.
Is there anything out there like that?
Vendors are welcome to respond.
TIA,
Julie Collins
Dir, Immunohistochemistry
Univ. of Kansas Med. Center
Kansas City, KS
----------------------------------------------------------------------
Date: 25 Oct 2001 14:35:25 -0500
From: "Weems, Joyce"
Subject: FW: OT - Memo
Thank goodness for our sense of humor.......
Memo from the office of the President
> >
> > Date: 14 September 2001
> > To: Albert Gore
> > Dear Al:
> > We found some more votes. You won. When do you want
to take over?
> > Sincerely,
> > George W. Bush
----------------------------------------------------------------------
Date: 25 Oct 2001 14:35:59 -0500
From: "Weems, Joyce"
Subject: RE: Personnel Shortage
Gracious! Can I sign up. I'll learn to do Cytology!!
Joyce Weems
Pathology Manager
Saint Joseph's Hospital of Atlanta
-----Original Message-----
From: Bruce Gapinski
[SMTP:BGapinski@pathgroup.com]
Sent: Thursday, October 25, 2001 10:58 AM
To: Weems, Joyce
Cc: 'HistoNet@Pathology.swmed.edu'
Subject: RE: Personnel Shortage
Does this mean another raise for Cytology? Our
Cytology department
got a 30%
raise last year! We got 2%. I'm sick of it! <--
sorry, venting.
Respectfully,
Bruce Gapinski HT(ASCP)
-----Original Message-----
From: Weems, Joyce
[mailto:JWEEMS@sjha.org]
Sent: Thursday, October 25, 2001
6:40 AM
To: 'Histonet'
Subject: FW: Personnel
Shortage
This is an email I received from my
Director
regarding the
Congressional
Bill. Perhaps it will help clarify
the email sent by
Glenda
Hoye
.
The bill does specifically mention
Histology and
Cytology.
Send your response to
ElissaP@ascls.org
or call or fax as below.
We all need to speak out! Next year
is a BIG
election year.
Use that to our
advantage. We need to speak directly
to our Senators
and
Representatives
also - emails, voice mails, etc.
I belong to the Advocacy Committee -
I will forward
things
along as I see
relevance. (Excuse us, Dear Friends
outside the
USA!)
God bless us everyone!
Joyce Weems
Pathology Manager
Saint Joseph's Hospital of Atlanta
Subject: Personnel Shortage
Hi Everyone,
As this Congress winds down its
work, there is some
concern
that H.R. 1948
will not be enacted. In addition,
since there never
was a
Senate bill, it
is unlikely that any of it would be
implemented. In
meetings that we
(ASCLS) have been having with the
College of
American
Pathologists, we have
discussed what it would really take
to fix the
shortage.
The College's
contacts on the Hill have indicated
that they
believe that
there is a
shortage but that loan forgiveness
programs aren't
really
going to fix the
problem. David Fowler reported that
Secretary
Thompson
(HHS), as part of a
video conference on anthrax, stated
four different
times
about the
difficulty the labs were having with
keeping up with
the
amount of work and
stated that this was a concern that
was brought to
his
attention and needs
to be addressed. He also stated that
he needs some
direction
about the best
way to help the labs. As David
said, this is the
time for
us to tell folks
what is needed for the short and
long term to make
sure that
the public gets
the level of testing and quality
that they deserve.
CAP
agrees and is
trying to get a meeting with the
Secretary for the
two
organizations (and
other organizations that might want
to attend). CAP
is
asking us to come up
with some really effective
strategies to fix this
shortage
so that CAP can
endorse them and we can all be
sending the same
message.
Therefore, we need
to do some brainstorming. We know
that salary and
recognition are two
crucial contributors to the
shortage. But are there
other
things? And what
is the answer?
So, please talk among yourselves,
share this email
with as
many folks as you
would like but get back to David
and/or I with:
a. what is contributing to the
shortage (with some
prioritization of the
list)
b. what will really fix it?
(including money - but
money
where - to just
the hospitals, to education
programs, to ? where)
Please get back to us as soon as
possible but no
later than
Oct 31. Thanks!
Elissa Passiment
ASCLS Executive Vice President
301.657.2768, X3015
301.657.2909 (fax)
7910 Woodmont Ave, #530
Bethesda, MD 20814-3015
----------------------------------------------------------------------
Date: 25 Oct 2001 14:36:32 -0500
From: "Nava, Josefa"
Subject:
----------------------------------------------------------------------
Date: 25 Oct 2001 14:36:57 -0500
From: Patti Loykasek
Subject: Re: MyoD1
We obtain the best results with this antibody using it with
a citrate
pressure cook retrieval at 1:25 overnight incubation in the
frig, LSAB+
detection. We use a fetal limb as a positive control.
Whenever testing a new
antibody we do a variety of pretreatments and titers. Hope
this helps. If
you have any further questions, please contact me.
Patti Loykasek
Phenopath Laboratories
Seattle, WA
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Date: 25 Oct 2001 14:37:17 -0500
From: Luis Chiriboga
Subject: Looking for a supplier of........
TIA-1
Hi fellow histonetters
Does any know where I can get find TIA-1.
Thanks in advance
LC
- --------------------------------------
----------------------------------------------------------------------
Date: 25 Oct 2001 14:37:38 -0500
From: Richard Cartun
Subject: Looking for Rosalba Zumbo
Would Rosalba Zumbo (Technical Officer/Scientific at the
Institute of Forensic
Medicine in Sydney, Australia) please contact me. Thanks!
R. Cartun
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Date: 25 Oct 2001 14:37:59 -0500
From: "Sebree Linda A."
Subject: RE: MyoD1
Hi Linda,
I can tell how we used to stain for MyoD1 (we have since
switched to
Myogenin). We used Myo D1 from NeoMarkers at 1:30 for 32"
incubation on
Ventana automated instruments. Our pretreatment was 4" in
Biocare's Nuclear
DeCloaker in Biocare's DeCloaking chamber. We further
amplified the primary
antibody signal with Ventana's amplification kit (rabbit X
mouse IgG
followed by mouse X rabbit IgG).
Hope some of this is of use to you,
Linda A. Sebree, HT(ASCP)
University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
A4/204-2472
600 Highland Ave.
Madison, WI 53792-2472
(608)265-6596
FAX: (608)262-7174
- -----Original Message-----
From: Lynde77627@aol.com [mailto:Lynde77627@aol.com]
Sent: Thursday, October 25, 2001 12:05 PM
To: histonet@pathology.swmed.edu
Subject: MyoD1
I saw a note several weeks ago requesting help with MyoD1
but never saw any
responses. I am now in the process of attempting to get it
to work. If
anyone has any insite to this antibody I sure would
appreciate it. I am
using FFPE tissue and NovaCastra antibody.
Thanks in advance.
Linda
----------------------------------------------------------------------
Date: 25 Oct 2001 15:59:33 -0500
From: Dana Settembre
Subject: Re: MyoD1
Hello Linda
I use the monoclonal antibody from Dako code # M3512 at a
dilution of 1:25. I
also use their Target Retreval System (TRS) in a steamer for
about 40 minutes.
See if you could try your antibody with Dako's TRS in a
steamer. Good Luck.
Dana Settembre
Immunohistochemistry Lab
University Hospital - UMDNJ
Newark, NJ
>>> 10/25/01 01:04PM >>>
I saw a note several weeks ago requesting help with MyoD1
but never saw any
responses. I am now in the process of attempting to get it
to work. If
anyone has any insite to this antibody I sure would
appreciate it. I am using
FFPE tissue and NovaCastra antibody.
Thanks in advance.
Linda
----------------------------------------------------------------------
Date: 25 Oct 2001 16:00:15 -0500
From: "Allen, Steven"
Subject: Hansel's stain
Hello,
We are looking for a procedure for Hansel's stain. Has
anybody done this in
their lab, and do you have any hints/tips for this
stain?Thanks in advance!
Steve Allen
Lovelace Respiratory Research Institute
2425 Ridgecrest SE Albuquerque, NM 87108
(505)348-9159
----------------------------------------------------------------------
Date: 25 Oct 2001 16:01:25 -0500
From: "Sebree Linda A."
Subject: RE: microtome help
I too have fond memories of using an old black AO; and no I
don't think
there is anything like it out there (or as good!). Our lab
is currently
using an Olympus Cut 4060 which does a pretty decent job on
most specimens
but I sometimes have problems with extremely dense tissue
thick and thinning
even after tightening everything, chilling the block and
using a new blade.
I think that one can buy a workhorse type microtome that can
handle all
tissues adequately but not necessarily elegantly or
something like an
Olympus that produces beautiful thin sections of things like
lymph nodes but
can't always stand up to all tissue types.
This is of course only my opinion after 20+ years in the
field and I do not
have experience with everything on the market these days.
As for price, I wouldn't expect to get a really good, new
microtome cheaply.
We paid $7,500 seven years ago for our instrument.
Linda A. Sebree, HT(ASCP)
University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
A4/204-2472
600 Highland Ave.
Madison, WI 53792-2472
(608)265-6596
FAX: (608)262-7174
- -----Original Message-----
From: Julie Collins [mailto:jcollins@kumc.edu]
Sent: Thursday, October 25, 2001 12:51 PM
To: HistoNet@pathology.swmed.edu
Subject: microtome help
Can anyone tell me where I can get a decent inexpensive, new
microtome? I
haven't purchased one in 10 years and have no idea where to
start looking.
I don't need anything fancy. My favorite microtome I had
was the old black
AO. Is there anything out there like that?
Vendors are welcome to respond.
TIA,
Julie Collins
Dir, Immunohistochemistry
Univ. of Kansas Med. Center
Kansas City, KS
----------------------------------------------------------------------
Date: 25 Oct 2001 16:01:48 -0500
From: Jan Shivers
Subject: Fw: OT
And a huge thank you, also, to Prime Minister Tony Blair,
who has been so
overtly supportive of the US. I'm sure there are citizens
of the UK who
wish he were not so vocal; he could be taking a risk at
putting his own
country in harm's way as well. It's at times like these
when one realizes
that we are all citizens of the world, not just of our own
little corner.
Believe me, we Americans are SO grateful for the kind words
and supportive
actions of others around the globe. It has meant more than
you can know.
We hope that you friends from other countries stay safe in
these crazy
times.
Jan Shivers
Univ. of Minnesota, USA
- ----- Original Message -----
From: "Connie McManus"
To: "Barry Rittman" ;
"histology"
Sent: Thursday, October 25, 2001 7:34 AM
Subject: Re: OT
> Barry,
> Thank you for sharing this! I am a born and bred
American, lived in the
> wild west most of my life, but I have a great love for the
Brits, too, and
> am DEEPLY greatful for their support of our President in
these times...
> THANKS TO THE UK!! *G*
>
> Up with the Union Jack!
> Connie McManus
>
> At 06:59 AM 10/25/01 -0500, Barry Rittman wrote:
> >A former student who is in the US navy came to visit us.
He told us of
> >an incident about three weeks ago when his ship was being
passed by a
> >British destroyer.
> >The sailors on the destroyer appeared on deck in full
dress uniforms,
> >saluted the US ship and ran up the Stars and Stripes.
> >While I was born England I have come to love and respect
America and
> >what both countries stand for.
> >I found this gesture very touching and although I cannot
thank the
> >sailors directly I would like to thank Britons as a whole
for their
> >support in these difficult times.
> >Thank you.
> >Barry
> >
> >
> >
>
> Veterinary Diagnostics Lab
> Utah State University
> Logan, UT
> USA
> (435) 797-1891
> fax (435) 797-2805
>
>
----------------------------------------------------------------------
Date: 25 Oct 2001 16:22:15 -0500
From: Connie McManus
Subject: Re: FW: OT - Memo
My, oh my how we have change in such a short time! This is
great, thnkx
for sharing.
ConnieM
At 02:01 PM 10/25/01 -0400, Weems, Joyce wrote:
>Thank goodness for our sense of humor.......
>
>
>Memo from the office of the President
>> >
>> > Date: 14 September 2001
>> > To: Albert Gore
>
>> > Dear Al:
>
>> > We found some more votes. You won. When do you want
to take over?
>
>> > Sincerely,
>> > George W. Bush
>
>
Veterinary Diagnostics Lab
Utah State University
Logan, UT
USA
(435) 797-1891
fax (435) 797-2805
----------------------------------------------------------------------
Date: 25 Oct 2001 16:22:37 -0500
From: Connie McManus
Subject: Re: anthrax endospore staining
At 10:16 AM 10/24/01 -0600, Gayle Callis wrote:
>It is gram positive rods, but not having done actual
infected tissue, don't
>know how hard it is to find bacillus. One microbiologist
here showed a
>photo of bacillus with spores, stained with a polychrome
methylene blue -
>wonderful staining of the spores.
Did this actually stain the endospores??? usually, these
are impervious to
dyes and apear as clear, refractile bodies within the
vegetative cell.
however, the only method for staining them than I know of
uses malchite
green. This sounds really cool, Gayle. I'd love to see
it... wish I could.
Connie M.
>
>Check out some microbiology texts and pathology books
describing
>pathogenesis of disease, maybe these will have some photos
of what you want
>to see. Let us know what you find, it would be educational
for all of us.
>Hopefuly our CDC Histonetters will lead us in the right
direction.
>
>
>Gayle Callis
>MT,HT,HTL(ASCP)
>Histopathology Supervisor
>Veterinary Molecular Biology - Marsh Lab
>Montana State University - Bozeman
>19th and Lincoln St
>Bozeman MT 59717-3610
>
>406 994-6367
>406 994-4303 (FAX)
>
>
>
Veterinary Diagnostics Lab
Utah State University
Logan, UT
USA
(435) 797-1891
fax (435) 797-2805
----------------------------------------------------------------------
Date: 25 Oct 2001 16:23:04 -0500
From: Tom.Wells@nshr.hnet.bc.ca
Subject: Re: IHC 101
A great book to start with is "Introduction to
Immunocytochemistry", 2nd
edition, J.M. Polak and S. Van Noorden, Bios Scientific
Publishers. Tom
Wells
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A great book to start
with is
"Introduction to Immunocytochemistry", 2nd
edition, J.M. Polak and
S. Van Noorden, Bios Scientific Publishers. Tom Wells
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----------------------------------------------------------------------
Date: 25 Oct 2001 17:07:53 -0500
From: Richard Cartun
Subject: Re: Looking for a supplier of........
We use ImmunoTech's anti-TIA mAb.
R. Cartun
>>> Luis Chiriboga 10/25/01
03:15PM >>>
TIA-1
Hi fellow histonetters
Does any know where I can get find TIA-1.
Thanks in advance
LC
- --------------------------------------
----------------------------------------------------------------------
Date: 25 Oct 2001 17:52:17 -0500
From: GREYTRUNK@aol.com
Subject: Re: microtome help
I just bought a brand new leica for under 10k and I love it,
wouldn't trade
it for anything.
Roxanne Soto HT (ASCP)
Wilson Medical Center
Wilson, NC
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I just
bought a brand new leica for under 10k and I love it,
wouldn't trade it for
anything.
Roxanne Soto HT (ASCP)
Wilson Medical Center
Wilson, NC