RE: unsubscribe
| From: | "Ryan, Genoula K DHCS-Ft Belvoir" |
unsubscribe
-----Original Message-----
From: HistoNet Server [mailto:histonet@pathology.swmed.edu]
Sent: Saturday, October 19, 2002 12:58 AM
To: HistoNet Server
Subject: Daily Digest
----------------------------------------------------------------------
Date: 18 Oct 2002 00:30:40 -0500
From: "J. A. Kiernan"
Subject: Re: Paraffin temp for immuno's
Ross Stapf (or was it Patsy Ruegg) wrote:
> On searching ... why heat in paraffin ... might be more
> detrimental than heat in antigen retrieval solution.
> ... can't find if anyone came to a conclusion that this was true ...
> I'm sure someone has studied this in order to come up with this 56
The cited temperature 56C must mean that this is another urban
laboratory
legend! Anyone studying effects of wax temperature would
investigate
steps such as 50-55-60. An investigation with a precision of one
degree would need 20 lots of trials to cover the range, and what
agency would fund such a study?
Several anecdotal Histonet replies have reported exactly the
opposite - that temperatures well above the mid-50s does more
good than harm when it comes to detecting antigens
immunohistochemically.
These informal reports, based on firt-hand experience, carry much
more
weight than "someone said that..." or a photocopy of some piece
of
paper found in a cardboard box.
There is also a Common Wisdom of Immunohistochemistry that says
coagulation of proteins by heat or coagulant fixatives liberates
the epitopes of insolubilized protein molecules.
If the routine fixative everywhere was a simple alcohol-acetic
mixture, would there be any of these problems with masked
antigens?
- --
- -------------------------
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London, Canada N6A 5C1
kiernan@uwo.ca
http://publish.uwo.ca/~jkiernan/
----------------------------------------------------------------------
Date: 18 Oct 2002 01:00:36 -0500
From: jay cheung
Subject: silver stain and flurojade staining
Hi, everyone:
I am currently running silver staining and fluoro-jade staining for
detecting neuronal degeneration in rat brain fixed and fresh frozen
sections. I never ran these procedures before and the results I got
from
the first trial showed many non specific staining not limited to
degenerating neurons. Also, it seems like the PFA fixed tissue got
more
silver stain than the fresh post-fixed tissue. Can someone tell me the
mechanism of action of these staining procedures and what are the tricks
about
these procedures. I did some research myself and so far have no luck
finding out the answers for these questions. I used FD silver stain
kit
to do the silver stain. Thank you so much for your help!
Jay
Choose an Internet access plan right
for
you -- try MSN! Click Here
----------------------------------------------------------------------
Date: 18 Oct 2002 02:00:37 -0500
From: jim
Subject: Thickness
Dear Histonetters
has anyone had experience with myelin stains on thick sections i.e. 30-50
microns.
Jim
----------------------------------------------------------------------
Date: 18 Oct 2002 03:45:29 -0500
From: naveeda@akunet.org
Subject: lymph node biopsies
dear histo netters
any one of you can help me in our lab we are having problems with lymph node
biopsies. the problem is that that some of our lymph node biopsies are
cracked
and some become hard. we receive samples from our hospital as well as from
other hospitals the samples which we receive from ouside of our hospital
have
pH of buffered formalin sometimes around 6.0. does this has any effect on
these biopsies.
if any one can provide me lymph node biopsy processing and fixing protocol
thanks
naveeda
- --
Naveeda Arshad
Pathology Department
Aga Khan university Hospital
Karachi
Pakistan
- --
----------------------------------------------------------------------
Date: 18 Oct 2002 04:00:53 -0500
From: Chris van der Loos
Subject: Glucose Oxidase
-
----------------------------------------------------------------------------
----------------------------------------------------------------------
Luis Chiriboga wrote:
Date: 17 Oct 2002 11:07:15 -0500
From: Luis Chiriboga
Subject: Glucose Oxidase
Hi all
Has anyone ever tried to detect Glucose oxidase enzyme activity directly in
FF-PE tissues???
Any suggestions would be helpful
Dear all,
The application of GOX from Aspergillus niger as marker enzyme is
theoretically ideal since the GOX enzyme is not present in mammals. During
the 70-80ties GOX as enzymatic marker was even commercially available!
However, in my hands and that of many others background of unknown origin
occurred. As far as I am aware of nobody is using GOX anymore.
For staining protocol see: Campbell and Bhatnagar, J Histochem Cytochem
24:448 (1976) and Clark et al J Histochem Cytochem 30:27 (1982)
After removing some dust the following protocol appeared:
Tris-HCl buffer (0.1 M, pH8.3) 5.0 ml
b-D(+) glucose (Sigma G5250) 67 mg
NBT (Sigma N6876) 5 mg/ml 1.34 ml
pPMS (Sigma P9625) 1mg/2 ml dist. water 0.34 ml (keep in dark!)
dist. water 3.33 ml
Incubate at 37#161#C in the dark for 30-60 min
wash with distilled water
mount aqueously
Success,
Chris
-
----------------------------------------------------------------------------
--------------------------------------------------------------
----------------------------------------------------------------------
Date: 18 Oct 2002 04:31:01 -0500
From: Chris van der Loos
Subject: RE: in situ hybridization course
-
----------------------------------------------------------------------------
--------------------------------------------
Philip Wan wrote:
Date: 17 Oct 2002 16:30:25 -0500
From: "Wan, Philip"
Subject: in-situ hybridization
Can anyone recommend any professional short training course classes for
learning fluorescent in situ hybridization techniques? Particularly in
relation to identifying gene copy number and chromosomal location.
I figured since immunoassays in histology frequently utilize in situ
hybridization techniques, someone here might just know of a course that
would be of use to me.
Thank you for your time,
Phil Wan
Dear Phil,
You may find a good 1-week course in spring 2003 in Europe.
Please check www.hbtc.nl. Move to "English", than click "in situ
hybridization". You will find all info up here.
Chris
_---------------------------------------------------------------------------
---------------------------------------
----------------------------------------------------------------------
Date: 18 Oct 2002 08:00:45 -0500
From: PMarcum
Subject: RE: Paraffin temp for immuno's
One issue for paraffin manufacturers is the lower the melt point of wax the
softer the wax and the more additive we have to add. Most labs prefer a
firmer paraffin without the stickiness of high additive thus we look at the
55-60C melt point paraffins as the base. The urban legend may have as much
to do with what we like to cut as it does with reality of IHC. The lower
melt points require much colder cutting conditions for excellent ribbons or
a great deal of compression from a warming block. Pam Marcum
> -----Original Message-----
> From: J. A. Kiernan [mailto:jkiernan@uwo.ca]
> Sent: Friday, October 18, 2002 1:19 AM
> To: histonet@pathology.swmed.edu
> Subject: Re: Paraffin temp for immuno's
>
>
> Ross Stapf (or was it Patsy Ruegg) wrote:
> > On searching ... why heat in paraffin ... might be more
> > detrimental than heat in antigen retrieval solution.
> > ... can't find if anyone came to a conclusion that this was true ...
> > I'm sure someone has studied this in order to come up with this 56
>
> The cited temperature 56C must mean that this is another urban
> laboratory
> legend! Anyone studying effects of wax temperature would
> investigate
> steps such as 50-55-60. An investigation with a precision of one
> degree would need 20 lots of trials to cover the range, and what
> agency would fund such a study?
>
> Several anecdotal Histonet replies have reported exactly the
> opposite - that temperatures well above the mid-50s does more
> good than harm when it comes to detecting antigens
> immunohistochemically.
> These informal reports, based on firt-hand experience, carry much
> more
> weight than "someone said that..." or a photocopy of some piece
> of
> paper found in a cardboard box.
>
> There is also a Common Wisdom of Immunohistochemistry that says
> coagulation of proteins by heat or coagulant fixatives liberates
> the epitopes of insolubilized protein molecules.
>
> If the routine fixative everywhere was a simple alcohol-acetic
> mixture, would there be any of these problems with masked
> antigens?
>
> --
> -------------------------
> John A. Kiernan
> Department of Anatomy and Cell Biology
> The University of Western Ontario
> London, Canada N6A 5C1
> kiernan@uwo.ca
> http://publish.uwo.ca/~jkiernan/
>
>
----------------------------------------------------------------------
Date: 18 Oct 2002 08:01:06 -0500
From: "Johnston, Kathy"
Subject: RE: Questions about staining solutions.
We change our solutions for M. Trichromes every two weeks, and they are used
daily with slide counts ranging from 5 to 50 or more per day.
Kathy
- -----Original Message-----
From: Coleman, Jeffrey [mailto:jeffrey.coleman@canji.com]
Sent: Thursday, October 17, 2002 1:52 PM
To: HistoNet Server (E-mail)
Subject: Questions about staining solutions.
I'm trying to set up a schedule for changing the stains we use in our Mikrom
Robot Stainer. However, I'm having a problem figuring out how long some of
the stains are usable.
For H&E stains, I've found some literature suggesting the hematoxylin can be
used for up to 6 months, some say 2 months, others say depending on the pH.
Does anyone know what the pH for a good hematoxlin stain should be? How
often should the hematoxylin be changed?
We also use the Robot Stainer for Masson TriChrome stains. We've got a
changing schedule for all the solutions except Biebrich Scarlet/Acid Fuchsin
solution and Aniline Blue Stain solution. Any suggestions as to how often
these solutions should be changed?
Thanks in advance!
Jeff Coleman
Pharmacology, Canji
Schering-Plough Co.
***************************************************************
This message and any attachments is solely for the intended recipient. If
you are not the intended recipient, disclosure, copying, use, or
distribution of the information included in this message is prohibited --
please immediately and permanently delete this message.
----------------------------------------------------------------------
Date: 18 Oct 2002 08:31:13 -0500
From: MTitford@aol.com
Subject: TB/anecdotal evidence
I would like to add my two bits worth to the TB discussion. Anecdotal
evidence
is supposed to have no place in science, but here goes!
TB is endemic here along the Gulf of Mexico coast. In the last year we
have
had two "outbreaks" of TB exposures.One was a foriegn student who died
suddenly of TB and exposed other students. The second a local commercial
fisherman who exposed others in his community. The health department is
still
working on the second case.
In the college case, the local health department skin tested everyone who
had been exposed. Those who were positive were X-rayed.(All at no cost).
Just
about everyone who had positive skin tests had negative X-rays. Those who
were
negative but were still worried because of exposure were offered the long
course of medication.
I get the impression that many convert for one reason or another, but few
get
the disease, because our standard of living is so much better now that when
TB
was a menace years ago.
People who grew up in England in the 50's may have been given BCG, a type
of
immunization against TB which I think is no longer given. BCG gives you an
automatic positive PPD which can put unknowing health care workers in a
frenzy
if they are unfamilier with BCG.
Mike Titford
Pathology
USA Medical Center
Mobile AL USA
----------------------------------------------------------------------
Date: 18 Oct 2002 08:46:03 -0500
From: DDittus787@aol.com
Subject: Re: Questions about staining solutions.
We change our stains weekly,basically we use slide volume rather than ph,
but
since the higher number of slides, the more carryover of alcohols or water
from washes, then obviously ph changes,we change certain alcohols and
xylenes
daily, others we rotate all of course having an effect on the h&e quality.we
also run a test h&e slide every morning, i suggest you may want to do this
with your trichromes and can possibly determine maximum number of slides
before quality gives out. just one more histotechs opinion.
dana
----------------------------------------------------------------------
Date: 18 Oct 2002 09:01:04 -0500
From: silvia.2.pasquetto@gsk.com
Subject: Courses..........
Does anybody know if there are courses about electron microscopy
techniques that I, as a beginner, can't miss?
"Chris van der Loos"
18-Oct-2002 11:19
To: histonet
cc:
Subject: RE: in situ hybridization course
-
----------------------------------------------------------------------------
--------------------------------------------
Philip Wan wrote:
Date: 17 Oct 2002 16:30:25 -0500
From: "Wan, Philip"
Subject: in-situ hybridization
Can anyone recommend any professional short training course classes for
learning fluorescent in situ hybridization techniques? Particularly in
relation to identifying gene copy number and chromosomal location.
I figured since immunoassays in histology frequently utilize in situ
hybridization techniques, someone here might just know of a course that
would be of use to me.
Thank you for your time,
Phil Wan
Dear Phil,
You may find a good 1-week course in spring 2003 in Europe.
Please check www.hbtc.nl. Move to "English", than click "in situ
hybridization". You will find all info up here.
Chris
_---------------------------------------------------------------------------
---------------------------------------
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contained in the following MIME Information.
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Does anybody know if there are courses about
electron microscopy techniques that I, as a beginner, can't miss?
|
| "Chris van der Loos"
<c.m.vanderloos@amc.uva.nl>
18-Oct-2002 11:19
|
To:
histonet
cc:
Subject:
RE: in situ hybridization course |
-----------------------------------------------------------------------
-------------------------------------------------
Philip Wan wrote:
Date: 17 Oct 2002 16:30:25 -0500
From: "Wan, Philip" <PWan@medarex.com>
Subject: in-situ hybridization
Can anyone recommend any professional short training course classes for
learning fluorescent in situ hybridization techniques? Particularly in
relation to identifying gene copy number and chromosomal location.
I figured since immunoassays in histology frequently utilize in situ
hybridization techniques, someone here might just know of a course that
would be of use to me.
Thank you for your time,
Phil Wan
Dear Phil,
You may find a good 1-week course in spring 2003 in Europe.
Please check www.hbtc.nl. Move to "English", than click "in
situ
hybridization". You will find all info up here.
Chris
_---------------------------------------------------------------------------
---------------------------------------
- --Boundary_(ID_21uhbHzEJ36HU82j/P0SMA)--
----------------------------------------------------------------------
Date: 18 Oct 2002 09:15:39 -0500
From: Garry Ashton
Subject: histology description
Dear all,
I am looking for books / references which best describe histology, the
subject and the job.
Do any of you histonetters know of any.
Thanks in advance.
Garry
PICR
UK
This email is confidential and intended solely for the use of the Person(s)
('the intended recipient') to whom it was addressed. Any views or opinions
presented are solely those of the author and do not necessarily represent
those of the Paterson Institute For Cancer Research or the Christie Hospital
NHS Trust. It may contain information that is privileged & confidential
within the meaning of applicable law. Accordingly any dissemination,
distribution, copying, or other use of this message, or any of its contents,
by any person other than the intended recipient may constitute a breach of
civil or criminal law and is strictly prohibited. If you are NOT the
intended recipient please contact the sender and dispose of this e-mail as
soon as possible.
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Dear
all,
I am looking for
books / references which best describe histology, the subject and the
job.
Do any of you
histonetters know of any.
Thanks in
advance.
Garry
PICR
UK
This email is confidential and intended solely
for the use of the Person(s) ('the intended recipient') to whom it was
addressed. Any views or opinions presented are solely those of the author
and
do not necessarily represent those of the Paterson Institute For Cancer
Research or the Christie Hospital NHS Trust. It may contain information that
is privileged & confidential within the meaning of applicable law.
Accordingly
any dissemination, distribution, copying, or other use of this message, or
any
of its contents, by any person other than the intended recipient may
constitute a breach of civil or criminal law and is strictly prohibited. If
you are NOT the intended recipient please contact the sender and dispose of
this e-mail as soon as possible.
- --Boundary_(ID_iIvu6Clv5CIukpuwD/ogKg)--
----------------------------------------------------------------------
Date: 18 Oct 2002 09:30:40 -0500
From: Cynthia Favara
Subject: RE: skull decalcification
Matt,
I have done fixed sucrose protected frozen sections and paraffin embedded
sections on neonatal mice up to 2 weeks of age without decalcification. I
have used a few strains of mice and found the skull cap to be thin enough to
section with little or no difficulty. I found careful dissection after
fixation to remove the lower jaw is necessary.
As usual Gayle gave a good protocol for decalcification if necessary.
Cynthia Favara
NIAID/NIH/RML/LPVD
903 South 4th Street
Hamilton, MT 59840
406-363-9317
- -----Original Message-----
From: Matt McElwee [mailto:MCELWEE@surgery.wisc.edu]
Sent: Thursday, October 17, 2002 8:15 AM
To: histonet@pathology.swmed.edu
Subject: skull decalcification
Hi everyone. Does anyone have a good way to decalcify mouse skulls that
doesn't shrink or otherwise alter the brain? We want to section mouse
heads from neonates through 2 weeks of age. I know that we'll have to
change the time that the brain is in the decalcifying solution,
depending on the age of the mouse, but if anyone can recommend a good
decalcifying solution or give any other advice, I'd really appreciate
it.
Thank you.
Matt
Matt McElwee
Research Specialist
Department of Surgery
K4/617 Clinical Science Center
600 Highland Avenue
Madison, WI 53792
(608) 263-7648
mcelwee@surgery.wisc.edu
----------------------------------------------------------------------
Date: 18 Oct 2002 09:30:57 -0500
From: "Salcedo, Rudy"
Subject: RE: frozen frog oocytes
I'm having the same problem here. I agree with the temp but still working
on it. Hope we get some response.
Rudy Salcedo
Research Assistant II
SCEH&M
Research Histopathology Core Lab
UTMB-Galveston
-----Original Message-----
From: Andrea Grantham [mailto:algranth@u.arizona.edu]
Sent: Wednesday, October 16, 2002 6:53 PM
To: histonet@pathology.swmed.edu
Subject: frozen frog oocytes
I'm trying to section Xenopus oocytes frozen in OCT with
poor results. Lots
of cracking and shattering. I'm figuring that temp plays a
roll here and
the little things are very "yolky".
Is there anybody out in histonetland who may have done some
sectioning of
frog oocytes or something similar? I actually found a
Xenopus histo book
but no suggestions for cryosectioning.
As is usually the case here, the labs bring the tissue
already frozen in
OCT. What can I suggest they do prior to or during freezing
the make the
sections cut better?
Does anybody know an optimal temp for cutting frog oocytes?
Thanks for any help.
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: 18 Oct 2002 09:37:54 -0500
From: Nick Bullough
Subject: RE: TB/anecdotal evidence
Hi Mike,
I'm only in my early 30's and was given BCG at school along with all my
fellow pupils, so BCG was still in use about 15 years ago and still is
today. In urban centres in the UK we have a major problem with immigrants
from the Indian subcontinent coming in to the country infected with TB and
passing it on to non-immunised people.
Best regards
Nick
Nick Bullough BSc(Hons) AIBMS
Scientist
Bioresources
Pharmagene plc
2 Orchard Road
Royston
Herts SG8 5HD
UK
nick.bullough@pharmagene.com
- -----Original Message-----
From: MTitford@aol.com [mailto:MTitford@aol.com]
Sent: 18 October 2002 14:24
To: Histonet@pathology.swmed.edu
Subject: TB/anecdotal evidence
I would like to add my two bits worth to the TB discussion. Anecdotal
evidence is supposed to have no place in science, but here goes!
TB is endemic here along the Gulf of Mexico coast. In the last year we
have had two "outbreaks" of TB exposures.One was a foriegn student who died
suddenly of TB and exposed other students. The second a local commercial
fisherman who exposed others in his community. The health department is
still working on the second case.
In the college case, the local health department skin tested everyone who
had been exposed. Those who were positive were X-rayed.(All at no cost).
Just about everyone who had positive skin tests had negative X-rays. Those
who were negative but were still worried because of exposure were offered
the long course of medication.
I get the impression that many convert for one reason or another, but few
get the disease, because our standard of living is so much better now that
when TB was a menace years ago.
People who grew up in England in the 50's may have been given BCG, a type
of immunization against TB which I think is no longer given. BCG gives you
an automatic positive PPD which can put unknowing health care workers in a
frenzy if they are unfamilier with BCG.
Mike Titford
Pathology
USA Medical Center
Mobile AL USA
****************************************************************************
**
This message and any files transmitted with it are intended for the
addressee
only and may contain information that is confidential or privileged.
Unauthorised use is strictly prohibited and may be unlawful. If you are not
the addressee, you should not read, copy,disclose or otherwise use this
message, except for the purpose of delivery to the addressee. If you
received
it in error please notify us immediately and then destroy it.
Further, Pharmagene makes every effort to keep its network free from viruses
including the scanning of incoming and outgoing mail. However, you do need
to
check this e-mail and any attachments to it for viruses as Pharmagene can
take
no responsibility for any computer virus that might be transferred by way of
this e-mail.
Pharmagene plc
2 Orchard Road,
Royston,
Hertfordshire,
SG8 5HD,
UK.
Registered in England & Wales under company number 03355618.
****************************************************************************
**
----------------------------------------------------------------------
Date: 18 Oct 2002 10:30:21 -0500
From: L.Wallez@chmouscron.be
Subject: ZINC Formalin
Any recipe of lab-made zinc formalin ?
Regards.
Luc Wallez, MD
----------------------------------------------------------------------
Date: 18 Oct 2002 10:30:41 -0500
From: Nick Bullough
Subject: RE: histology description
Garry,
Your best option would be to get in contact with the IBMS as they have
plenty information on the job. The website is www.ibms.org
(plus it all has a UK slant).
Best regards
Nick
Nick Bullough BSc(Hons) AIBMS
Scientist
Bioresources
Pharmagene plc
2 Orchard Road
Royston
Herts SG8 5HD
UK
nick.bullough@pharmagene.com
- -----Original Message-----
From: Garry Ashton [mailto:GAshton@PICR.man.ac.uk]
Sent: 18 October 2002 14:49
To: 'histonet'
Subject: histology description
Dear all,
I am looking for books / references which best describe histology, the
subject and the job.
Do any of you histonetters know of any.
Thanks in advance.
Garry
PICR
UK
This email is confidential and intended solely for the use of the Person(s)
('the intended recipient') to whom it was addressed. Any views or opinions
presented are solely those of the author and do not necessarily represent
those of the Paterson Institute For Cancer Research or the Christie Hospital
NHS Trust. It may contain information that is privileged & confidential
within the meaning of applicable law. Accordingly any dissemination,
distribution, copying, or other use of this message, or any of its contents,
by any person other than the intended recipient may constitute a breach of
civil or criminal law and is strictly prohibited. If you are NOT the
intended recipient please contact the sender and dispose of this e-mail as
soon as possible.
****************************************************************************
**
This message and any files transmitted with it are intended for the
addressee
only and may contain information that is confidential or privileged.
Unauthorised use is strictly prohibited and may be unlawful. If you are not
the addressee, you should not read, copy,disclose or otherwise use this
message, except for the purpose of delivery to the addressee. If you
received
it in error please notify us immediately and then destroy it.
Further, Pharmagene makes every effort to keep its network free from viruses
including the scanning of incoming and outgoing mail. However, you do need
to
check this e-mail and any attachments to it for viruses as Pharmagene can
take
no responsibility for any computer virus that might be transferred by way of
this e-mail.
Pharmagene plc
2 Orchard Road,
Royston,
Hertfordshire,
SG8 5HD,
UK.
Registered in England & Wales under company number 03355618.
****************************************************************************
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Garry,
Your best option would be to get in contact with
the
IBMS as they have plenty information on the job. The website is www.ibms.org (plus it all has a UK
slant).
Best regards
Nick
Nick Bullough BSc(Hons) AIBMS
Scientist
Bioresources
Pharmagene
plc
2 Orchard
Road
Royston
Herts
SG8
5HD
UK
nick.bullough@pharmagene.com
Dear
all,
I am looking
for
books / references which best describe histology, the subject and the
job.
Do any of you
histonetters know of any.
Thanks in
advance.
Garry
PICR
UK
This email is confidential and intended solely
for
the use of the Person(s) ('the intended recipient') to whom it was
addressed.
Any views or opinions presented are solely those of the author and do not
necessarily represent those of the Paterson Institute For Cancer Research
or
the Christie Hospital NHS Trust. It may contain information that is
privileged
& confidential within the meaning of applicable law. Accordingly any
dissemination, distribution, copying, or other use of this message, or any
of
its contents, by any person other than the intended recipient may
constitute
a
breach of civil or criminal law and is strictly prohibited. If you are NOT
the
intended recipient please contact the sender and dispose of this e-mail as
soon as possible.
****************************************************************************
**
This message and any files transmitted with it are intended for the
addressee
only and may contain information that is confidential or privileged.
Unauthorised use is strictly prohibited and may be unlawful. If you are not
the addressee, you should not read, copy,disclose or otherwise use this
message, except for the purpose of delivery to the addressee. If you
received
it in error please notify us immediately and then destroy it.
Further, Pharmagene makes every effort to keep its network free from viruses
including the scanning of incoming and outgoing mail. However, you do need
to
check this e-mail and any attachments to it for viruses as Pharmagene can
take
no responsibility for any computer virus that might be transferred by way of
this e-mail.
Pharmagene plc
2 Orchard Road,
Royston,
Hertfordshire,
SG8 5HD,
UK.
Registered in England & Wales under company number 03355618.
****************************************************************************
**
Here
are a couple websites that have basic info
Dear
all,
I am looking
for
books / references which best describe histology, the subject and the
job.
Do any of you
histonetters know of any.
Thanks in
advance.
Garry
PICR
UK
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----------------------------------------------------------------------
Date: 18 Oct 2002 11:46:14 -0500
From: "Willis, Donna"
Subject: Thymidylate Synthase Antibody
Does anyone in Histoland have a procedure for Thymidylate Synthase Antibody.
We have both DAKO and Ventana stainers and need help with this antibody. We
purchased the antibody from Chemicon.
Thanks,
Donna Willis
Histology Lab Manager
Harris Methodist Fort Worth, Tx
----------------------------------------------------------------------
Date: 18 Oct 2002 13:01:02 -0500
From: Eman Namati
Subject: Lung sectioning and imaging
I was wondering if anyone had embedded whole human or sheep lung? in MMA or
parafin?
and also if anyone had performed serial sections on a whole mouse lung
before?
Eman Namati
Project Manager, Lung Image Database Consortium
Roy J. and Lucille A. Carver College of Medicine
University of Iowa
200 Hawkins Dr., 378 MRF
Iowa City, IA 52242
Tel: (319) 384-8331
Mob: (319) 936-5487
Fax: (319) 353-6406
----------------------------------------------------------------------
Date: 18 Oct 2002 14:15:08 -0500
From: Eman Namati
Subject: Re: Lung sectioning and Imaing
I would like to get serial sections of an entire mouse lung. 3 micron
thickness every 30 microns.
For the human lung i would like to embedd the lung in half inch slabs of
200x150mm, so i can section it using a polycut microtome. I will be imaging
these off the block of remaining tissue using a stereoscope placed above the
microtome on a xyz gantry which will increase the field of view by acquiring
multple shots and creating one high res image of the entire section.
I would like to know if accurate serial sections through out an entire mouse
lung using conventional methods is possible or not? and has anyone tried
embedding whole human or sheep lung?
Eman Namati
Project Manager, Lung Image Database Consortium
Roy J. and Lucille A. Carver College of Medicine
University of Iowa
200 Hawkins Dr., 378 MRF
Iowa City, IA 52242
Tel: (319) 384-8331
Mob: (319) 936-5487
Fax: (319) 353-6406
----------------------------------------------------------------------
Date: 18 Oct 2002 14:30:46 -0500
From: Histo-Scientific Research Laboratories
Subject: Re: Lung sectioning and imaging
Eman,
We have done serial sectioning on whole mouse lungs and serial sections for
specific lung lobes of rats. If you have any questions regarding this
procedure feel free to give me a call directly.
Sincerely,
Tom Galati
Laboratory Director
HSRL- A GLP Compliant Laboratory
137 S. Main Street
Woodstock, VA 22664
(540)459-8211
fax: (540)459-8217
www.hsrl.org
tomgalati@hsrl.org
- ----- Original Message -----
From: "Eman Namati"
To: "HistoNet Server"
Sent: Friday, October 18, 2002 1:41 PM
Subject: Lung sectioning and imaging
> I was wondering if anyone had embedded whole human or sheep lung? in MMA
or
> parafin?
> and also if anyone had performed serial sections on a whole mouse lung
> before?
>
>
>
> Eman Namati
> Project Manager, Lung Image Database Consortium
> Roy J. and Lucille A. Carver College of Medicine
> University of Iowa
> 200 Hawkins Dr., 378 MRF
> Iowa City, IA 52242
> Tel: (319) 384-8331
> Mob: (319) 936-5487
> Fax: (319) 353-6406
>
>
>
----------------------------------------------------------------------
Date: 18 Oct 2002 14:31:11 -0500
From: Ross Stapf
Subject: Re: Paraffin temp for immuno's
Thank you to all who replied!
It seems like the good news is that I don't need to make any changes to my
paraffin.
As the reference was a 2002 book which was recently recomended here on
histonet, I thought maybe somebody knew something I hadn't heard of yet.
I think I have enough comments printed out to appease my Immuno Pathologist
if
he comes across this reference like I did.
Thank you all
Ross Stapf
>>> "J. A. Kiernan" 10/18/02 01:18AM >>>
Ross Stapf (or was it Patsy Ruegg) wrote:
> On searching ... why heat in paraffin ... might be more
> detrimental than heat in antigen retrieval solution.
> ... can't find if anyone came to a conclusion that this was true ...
> I'm sure someone has studied this in order to come up with this 56
The cited temperature 56C must mean that this is another urban
laboratory
legend! Anyone studying effects of wax temperature would
investigate
steps such as 50-55-60. An investigation with a precision of one
degree would need 20 lots of trials to cover the range, and what
agency would fund such a study?
Several anecdotal Histonet replies have reported exactly the
opposite - that temperatures well above the mid-50s does more
good than harm when it comes to detecting antigens
immunohistochemically.
These informal reports, based on firt-hand experience, carry much
more
weight than "someone said that..." or a photocopy of some piece
of
paper found in a cardboard box.
There is also a Common Wisdom of Immunohistochemistry that says
coagulation of proteins by heat or coagulant fixatives liberates
the epitopes of insolubilized protein molecules.
If the routine fixative everywhere was a simple alcohol-acetic
mixture, would there be any of these problems with masked
antigens?
- --
- -------------------------
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London, Canada N6A 5C1
kiernan@uwo.ca
http://publish.uwo.ca/~jkiernan/
----------------------------------------------------------------------
Date: 18 Oct 2002 15:15:38 -0500
From: hadams@mail.mdanderson.org
Subject: cryostat recommendation issues
Sorry, one other thing I forgot to include on cryostat prefs was service
quality, a very important component of any purchase. If anyone would care to
comment on frequency of repairs and the promptness of service when necessary
I
would be interested.
Hank Adams
Microscopy Core
Molecular Genetics Department
UT MD Anderson Cancer Center
Houston, Tx
----------------------------------------------------------------------
Date: 18 Oct 2002 15:45:57 -0500
From: LINDA GORMAN
Subject: tissue processors
We currently are using two 20+ year old VIPs and are looking to replace
one.We have no problem with purchasing another VIP,but we're trying to
be open-minded and give others a chance.We are now demoing a Ventana
Renasciance It appears to be doing a fine job but we are interested
in the long term.Have others used this instrument and found it to be
satisfactory?not Satisfactory?Please respond with your
experiences.Thanks.
Linda gorman
KU Med
----------------------------------------------------------------------
Date: 18 Oct 2002 15:46:19 -0500
From: pruegg@colobio.com
Subject: Re: TB Exposures
I am sure CDC keeps track of this.
Patsy
"Jasper, Thomas" wrote:
> My fellow histonetters,
>
> I would like to inquire about TB exposures. This would apply to
histotechs,
> PAs, pathologists, autopsy techs or any other lab personnel working in
> anatomic pathology. Specifically, how many with a prior negative PPD
> converted to a positive PPD after exposure to a known case? Demographics
> will factor into this inquiry so if you choose to respond it would be
> helpful to know where you're from. I do not wish to clog the net with
> responses, but others on this list may have interest in this data.
> Thanks!
>
> Thomas Jasper HT(ASCP)BAS
> Anatomic Pathology Coordinator
> SMDC Clinical Laboratory
> Duluth, MN
> tjasper@smdc.org
----------------------------------------------------------------------
Date: 18 Oct 2002 16:15:18 -0500
From: pruegg@colobio.com
Subject: Re: Paraffin temp for immuno's
As usual John enlightens us all. With the advent of HIER methods to
unmask proteins cross-linked by formalin our understanding of the tissue
biochemistry has changed, if we were not crosslinking the proteins with
formalin to begin with, we would not have to go through all the tissue
altering procedures (impossible to standardize from lab to lab) we do to
access the antigen site. I have for years advocated avoiding !0%NBF for
IHC studies. I have used zinc formalin myself for years and although it
is not formaldehyde free, it does not require near the retrieval 10%NBF
does. Enough for this soap box, I am afraid we have lost the battle to
change the 10% NBF choice of a standard fixative. We prefer too much to
look at archieval material which is already prepared this way, so we have
to jump through hoops trying to get at those antigens and reproduce IHC
results, hopefully in a standard way so the results can be compared from
shop to shop. Nobody ever said this was easy, did they???
Patsy
"J. A. Kiernan" wrote:
> Ross Stapf (or was it Patsy Ruegg) wrote:
> > On searching ... why heat in paraffin ... might be more
> > detrimental than heat in antigen retrieval solution.
> > ... can't find if anyone came to a conclusion that this was true ...
> > I'm sure someone has studied this in order to come up with this 56
>
> The cited temperature 56C must mean that this is another urban
> laboratory
> legend! Anyone studying effects of wax temperature would
> investigate
> steps such as 50-55-60. An investigation with a precision of one
> degree would need 20 lots of trials to cover the range, and what
> agency would fund such a study?
>
> Several anecdotal Histonet replies have reported exactly the
> opposite - that temperatures well above the mid-50s does more
> good than harm when it comes to detecting antigens
> immunohistochemically.
> These informal reports, based on firt-hand experience, carry much
> more
> weight than "someone said that..." or a photocopy of some piece
> of
> paper found in a cardboard box.
>
> There is also a Common Wisdom of Immunohistochemistry that says
> coagulation of proteins by heat or coagulant fixatives liberates
> the epitopes of insolubilized protein molecules.
>
> If the routine fixative everywhere was a simple alcohol-acetic
> mixture, would there be any of these problems with masked
> antigens?
>
> --
> -------------------------
> John A. Kiernan
> Department of Anatomy and Cell Biology
> The University of Western Ontario
> London, Canada N6A 5C1
> kiernan@uwo.ca
> http://publish.uwo.ca/~jkiernan/
----------------------------------------------------------------------
Date: 18 Oct 2002 18:00:58 -0500
From: Patti Loykasek
Subject: Re: Experience with CD4 and CD8 on paraffin sections?
> Here1s our protocol for CD4 on paraffin embedded. I have found it to be a
> astubborn1 antibody. Most of the time it works fine, but there are some
less
> than beautiful days. Hope this helps.
>
> Antibody: CD004
> Clone: 1F6
> Vendor: Novocastra
> Antigen Retrieval: Steam 40 minutes in EDTA, pH8
> Titer/Incubation: 1:10 for 40 minutes at room temperature
> Detection: DAKO Envision+, mouse for 30 minutes at room temperature
> Chromogen: DAB
>
> Patti Loykasek
> Phenopath Loykasek
> Seattle, WA
>
>
>
>
> Hello everyone,
> I am shopping for antibodies against human CD4 and CD8 to be used on
> paraffin-embedded tissue. I found 4 clones in NovoCastra, two against CD4
(1F6
> and 4B12) and two against CD8 (4B11 and 1A5). Does anyone have experience
with
> these clones? Thanks in advance,
> Emmanuel Maicas, MD in Moncton, Canada
>
>
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Re: Experience with CD4 and CD8 on paraffin sections?
Here’s our protocol for
CD4 on paraffin embedded. I have found it to be a ‘stubborn’
antibody. Most of the time it works fine, but there are some less than
beautiful days. Hope this helps.
Antibody: CD004
Clone: 1F6
Vendor: Novocastra
Antigen Retrieval: Steam 40 minutes in EDTA, pH8
Titer/Incubation: 1:10 for 40 minutes at room temperature
Detection: DAKO Envision+, mouse for 30 minutes at room temperature
Chromogen: DAB
Patti Loykasek
Phenopath Loykasek
Seattle, WA
Hello everyone,
I am shopping for antibodies against human CD4 and
CD8
to be used on paraffin-embedded tissue. I found 4 clones in NovoCastra, two
against CD4 (1F6 and 4B12) and two against CD8 (4B11 and 1A5). Does anyone
have experience with these clones? Thanks in advance,
Emmanuel Maicas, MD in Moncton, Canada
- --Boundary_(ID_wTTi6YwKkIj2VSnqsZdH+g)--
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