[Histonet] New Lab Design
Breeden, Sara
sbreeden <@t> nmda.nmsu.edu
Mon Aug 1 13:33:25 CDT 2005
Just a quick note to thank all of you that sent me your
ideas/comments/input on what to put into a new histo lab. These ideas
have been duly noted and will be part of My Grand Plan. There were some
meaty ideas and some real sparklers! Your help is invaluable -- and if
you've thought of anything else since last week, let me know. Thanks
again!
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
histonet-request <@t> lists.utsouthwestern.edu
Sent: Monday, August 01, 2005 11:02 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 21, Issue 1
Send Histonet mailing list submissions to
histonet <@t> lists.utsouthwestern.edu
To subscribe or unsubscribe via the World Wide Web, visit
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
or, via email, send a message with subject or body 'help' to
histonet-request <@t> lists.utsouthwestern.edu
You can reach the person managing the list at
histonet-owner <@t> lists.utsouthwestern.edu
When replying, please edit your Subject line so it is more specific than
"Re: Contents of Histonet digest..."
Today's Topics:
1. Excessive background staining with Millers elastic stain
(Lachlan Smith)
2. staining for iron in microglia (Sharon Cooperman)
3. Re: Excessive background staining with Millers elastic stain
(Paul Bradbury)
4. RE: New Lab Design (Rogerson Kemlo (ELHT) Pathology)
5. Biotinylating Antibodies (Orr, Rebecca)
6. Re: EGFR (DDittus787 <@t> aol.com)
7. (no subject) (Patsy Ruegg)
8. RE: (no subject) (Patsy Ruegg)
----------------------------------------------------------------------
Message: 1
Date: Mon, 1 Aug 2005 09:33:17 +0930
From: "Lachlan Smith" <lachlan.smith <@t> imvs.sa.gov.au>
Subject: [Histonet] Excessive background staining with Millers elastic
stain
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <000001c5962c$6b43cca0$e46b140a <@t> ITP36161>
Content-Type: text/plain; charset="iso-8859-1"
Dear Subscribers
I am staining cartilage using Millers elastic fibre stain, and while
elastic fibres are staining well, there is a lot of blue background
staining which makes thresholding for histoquantitation almost
impossible. My sections are 30 micron paraffin embedded. I am also
oxidising with 0.5% aqueous potassium permanaganate and bleaching with
2% oxalic acid.
Any suggestings for reducing this background staining would be greatly
appreciated.
Kind regards,
Lachlan Smith
Institute of Medical and Veterinary Science Adelaide, Australia
------------------------------
Message: 2
Date: Sun, 31 Jul 2005 21:39:40 -0400
From: Sharon Cooperman <scoop <@t> mail.nih.gov>
Subject: [Histonet] staining for iron in microglia
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <p06020420bf132ce1db09@[156.40.160.205]>
Content-Type: text/plain; charset="us-ascii" ; format="flowed"
Dear Histonetters,
I have a semi-scientific question: macrophages in almost all tissues
can be stained for iron with Perl's stain to give a blue color - is this
also true for microglia? If not, is there any other way to see how much
iron is in microglia (DAB enhanced Perl's or some other method)?
Thanks,
Sharon
--
Sharon Cooperman <scoop <@t> mail.nih.gov>
NIH, NICHD, CBMB 301.435-8417
Building 18T, room 101 301.402-0078 fax
Bethesda, MD 20892
------------------------------
Message: 3
Date: Sun, 31 Jul 2005 22:31:43 -0700
From: Paul Bradbury <histology.bc <@t> shaw.ca>
Subject: Re: [Histonet] Excessive background staining with Millers
elastic stain
To: Lachlan Smith <lachlan.smith <@t> imvs.sa.gov.au>, HistoNet Server
<histonet <@t> pathology.swmed.edu>
Message-ID: <42EDB3BF.3030304 <@t> shaw.ca>
Content-Type: text/plain; format=flowed; charset=us-ascii
Hi Lachlan,
I believe the problem you are having is due to the presence of very
similar reactive groups in both cartilage and elastic fibres, Elastin
(the protein ) is heavily sulphated with an abundance of disulphide
groups (hence the yellowish appearance of elastic fibres when viewed
unstained or even macroscopically). When you oxidize the sections in
potassium permanganate, you are converting the disulphides to sulphates,
the sulphates react with Miller's elastin stain. Miller's stain is
essentially as modification of aldeyde fuchsin which iis a well known
method for demonstrating sulphated groups in mucopolysaccharides, etc.
The extracellular matrix of cartilage consists of chondroitin sulphate
(plus a few other assorted bits of connective tissue). These groups will
react quite strongly with Miller's stain solution.
So unfortunately, I think you may well be out of luck as far as
improving the specificity of the staining reactions. The same low
specificity reactions occur with most other elastin staining methods
when they are used on cartilage, the reactive groups present in both
locations stain with the dye in use. I cannot think of a method that
stains only elastin and not cartilage matrix.
Thinner sections may give you a better chance of distinguishing one from
the other. Try sections at several thinner settings and see what the
outcome is. Maybe there is a thickness at which you can see sufficient
enough detail in the elastic fibres without the cartilage obscuring it.
Paul Bradbury,
Kamloops, BC
Canada
Lachlan Smith wrote:
>Dear Subscribers
>
>I am staining cartilage using Millers elastic fibre stain, and while
>elastic fibres are staining well, there is a lot of blue background
>staining which makes thresholding for histoquantitation almost
>impossible. My sections are 30 micron paraffin embedded. I am also
>oxidising with 0.5% aqueous potassium permanaganate and bleaching with
>2% oxalic acid.
>
>Any suggestings for reducing this background staining would be greatly
>appreciated.
>
>Kind regards,
>
>Lachlan Smith
>Institute of Medical and Veterinary Science Adelaide, Australia
>
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
------------------------------
Message: 4
Date: Mon, 1 Aug 2005 11:06:44 +0100
From: "Rogerson Kemlo (ELHT) Pathology" <Kemlo.Rogerson <@t> elht.nhs.uk>
Subject: RE: [Histonet] New Lab Design
To: "Breeden, Sara" <sbreeden <@t> nmda.nmsu.edu>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<53A22BBB01D6184EBF7A4DC18A021E9E5AD4C0 <@t> elht-exch1.xelht.nhs.uk>
Content-Type: text/plain; charset="us-ascii"
AFOS (or something similar), formalin extraction for grossing. Includes
a downdraught table and formalin making up facilities. AFOS formalin
extraction cabinets for stored samples in formalin.
Be careful over height of benches; some are for sitting down carrying
out work, others for cutting sections using a microtome. Nothing worse
than a microtome that's too high or too low. You need solvent extraction
hoods for your processing machines, a suitable flammable store with
antiflash lighting etc., The best lighting to get it that stuff they
sell that goes in Offices; it's expensive but gives you a good colour
temperature with few shadows.
If you can see, stop people asphyxiating on formalin and alcohol, stop
the place blowing up and have benches the right height that people can
work, nowt else much to do. Oh yes, coffee machine, clean area and a
Mars Bar vending machine. Ergo, all done!
-----Original Message-----
From: Breeden, Sara [mailto:sbreeden <@t> nmda.nmsu.edu]
Sent: 29 July 2005 17:27
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] New Lab Design
A new histology lab (in a new building) is in the works (yipee!!). I
have some definite ideas about what I'll need in my new digs, but I need
more opinions! Within the next couple weeks, I will be asked by the
architects for my input.
This is a State veterinary lab where I do "surgicals", necropsies,
standard special stains, with a rapidly expanding IHC workload. My total
workload was roughly 10,000 blocks last year and I expect it to grow by
5% each year. I expect to have roughly 500 sq ft to work with in this
new space. I work alone for three DVM pathologists.
What would you consider absolutely essential in a new histology lab?
What would you do differently if you'd had a chance? What kind of: bench
space and type of surface, ventilation, lighting, hoods, haz mat
storage, supply storage, windows? I only get once chance at this and I
want to make it count...I have 8 more years until retirement and I'd
rather not kick myself for missing something obvious.
Please reply directly to me. Thank you in advance!! I rely on 'netters
for great information.
Sally Breeden, HT(ASCP)
New Mexico Department of Agriculture
Veterinary Diagnostic Services
POBox 4700
Albuquerque, NM 87196
(505)841-2576
(505)841-2518 FAX
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 5
Date: Mon, 1 Aug 2005 07:26:53 -0500
From: "Orr, Rebecca" <ROrr <@t> enh.org>
Subject: [Histonet] Biotinylating Antibodies
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<A7FC3A4F98964A44B0B71B7B25EA6F9603AF425F <@t> EXCHANGE03.enhnet.org>
Content-Type: text/plain; charset="us-ascii"
Vernon,
I have been successful with a biotinylation kit from Biocare Medical.
It uses a different chemistry than the Vector kits.
I think the mopping reagent may be what makes the difference.
Their number is 800 799 9499 ask for Hari, he's the technical sales
manager.
Becky Orr, CLA HT (ASCP)
IHC Lead
Evanston Northwestern Healthcare
ph: 847-570-2771
------------------------------
Message: 6
Date: Mon, 1 Aug 2005 08:46:13 EDT
From: DDittus787 <@t> aol.com
Subject: Re: [Histonet] EGFR
To: JosefaNava <@t> texashealth.org, histonet <@t> pathology.swmed.edu
Message-ID: <9b.648c607f.301f7395 <@t> aol.com>
Content-Type: text/plain; charset="US-ASCII"
Josie:
Are you aware of the clone 31g7 and Allen Gowns ASCO 2005 poster. It
showed that 31g7 from Zymed was best in class and selected 10-15% more
patients.
dana
------------------------------
Message: 7
Date: Mon, 1 Aug 2005 10:21:23 -0600
From: "Patsy Ruegg" <pruegg <@t> ihctech.net>
Subject: [Histonet] (no subject)
To: "'Histonet'" <histonet <@t> pathology.swmed.edu>
Message-ID: <200508011621.j71GLFpf017202 <@t> chip.viawest.net>
Content-Type: text/plain; charset="us-ascii"
Does anyone know where I could get a service manual for an old IEC
cryostat?
Patsy
Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 216
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pruegg <@t> ihctech.net
web site www.ihctech.net <http://www.ihctech.net/>
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. 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.
------------------------------
Message: 8
Date: Mon, 1 Aug 2005 10:23:37 -0600
From: "Patsy Ruegg" <pruegg <@t> ihctech.net>
Subject: RE: [Histonet] (no subject)
To: <lachlan.smith <@t> imvs.sa.gov.au>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <200508011623.j71GNTpf017949 <@t> chip.viawest.net>
Content-Type: text/plain; charset="us-ascii"
We used picro sirrus stain for collagen fibers and viewed the collagen
bundles using polarized light filter for histomorphometry.
Patsy
Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 216
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pruegg <@t> ihctech.net
web site www.ihctech.net
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. 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.
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
lachlan.smith <@t> imvs.sa.gov.au
Sent: Sunday, July 31, 2005 3:18 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] (no subject)
Dear Subscribers
I am staining cartilage using Millers elastic fibre stain, and while
elastic
fibres are staining well, there is a lot of blue background staining
which
makes thresholding for histoquantitation almost impossible. My sections
are
30 micron paraffin embedded. I am also oxidising with 0.5% aqueous
potassium
permanaganate and bleaching with 2% oxalic acid.
Any suggestings for reducing this background staining would be greatly
appreciated.
Kind regards,
Lachlan Smith
Institute of Medical and Veterinary Science Adelaide, Australia
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
End of Histonet Digest, Vol 21, Issue 1
***************************************
More information about the Histonet
mailing list