[Histonet] RE: Histonet Digest, Vol 7, Issue 13
Dorothy.L.Webb <@t> HealthPartners.Com
Dorothy.L.Webb <@t> HealthPartners.Com
Thu Jun 24 14:56:51 CDT 2004
RE: Processing Toenails: We use NAIR for softening any toenails, etc. We
put it directly into the NAIR product for 2-4 hours and if it is a small
piece of nail, we cassette it(in filter paper or biopsy bags) and then place
it in the NAIR. Rinse it off with water and place in formalin for routine
processing.
-----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: Wednesday, June 09, 2004 12:07 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 7, Issue 13
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Today's Topics:
1. RE: Lymphocytes antibodies - Fixed tissue (Patsy Ruegg)
2. CORRECTIONLymphocytes antibodies - Fixed tissue (Patsy Ruegg)
3. RE: Double immunofluorescence with Alexa Fluor
(Pablo S?nchez Quinteiro)
4. JOB OPENING (Slydog5367 <@t> aol.com)
5. re: Automated Image analysis (Kevin Conway)
6. HPV results - probe vs. antibody (Kapoor, Sue)
7. Re: Lymphocytes antibodies - Fixed tissue (Drew Allan Roenneburg)
8. Re: HPV results - probe vs. antibody (DDittus787 <@t> aol.com)
9. Re: HPV results - probe vs. antibody (DDittus787 <@t> aol.com)
10. IHC Staining Basics (Julie Lee)
11. Reconditioned Leica Autostainer or Sikora Bach Stainer
(kevin williams)
12. Re: Microscopic Bubbles........by the hundreds & thousands
(Rosalba)
13. (no subject) (Lori Wilson)
14. Thanks for the helping find antibody (HCS)
15. renal carcinoma cells (Garry Ashton)
16. charge for cryostat use (Jill Songer)
17. blocking endogenous peroxidase when using ABC staining and
DAB (SMITH,REBEKAH FELICIA)
18. Re: blocking endogenous peroxidase when using ABC stainingand
DAB ( Katri Tuomala)
19. Re: charge for cryostat use (Andrea Grantham)
20. Help (Latitia.E.Floyd <@t> gsk.com)
21. Toe nails !!!!! (Linda Jones)
22. RE: Help (Elizabeth Chlipala)
23. Rat brain vascular casting using Mercox (Chris Bjornsson)
24. Re: Help (Gayle Callis)
----------------------------------------------------------------------
Message: 1
Date: Tue, 8 Jun 2004 11:08:13 -0600
From: "Patsy Ruegg" <pruegg <@t> colobio.com>
Subject: RE: [Histonet] Lymphocytes antibodies - Fixed tissue
To: "Marceau, Gabriel \(IAF\)\(LAVAL\)"
<Gabriel.Marceau <@t> inrs-iaf.uquebec.ca>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <NGBBIMHHKLGNCOCPDKBOEELMCHAA.pruegg <@t> colobio.com>
Content-Type: text/plain; charset="iso-8859-1"
Gabriel,
I use mouse monoclonal cd3 from Lab Vision for pan T cells on mouse tissue
ffpe, it should work great on frozen sections, unfortunately the t subsets
are not developed yet and work only on frozens so far.
Patsy
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Marceau,
Gabriel (IAF)(LAVAL)
Sent: Tuesday, June 08, 2004 8:53 AM
To: 'histonet <@t> lists.utsouthwestern.edu'
Subject: [Histonet] Lymphocytes antibodies - Fixed tissue
I Hi everyone,
I am looking for antibodies against lymphocytes (CD3 or other) that will
work on fixed tissue. Most of those comercially available are good for
cytometry but none is marked as usable on paraformaldehyde fixed tissue. If
someone tried one that is not specified for fixed tissue and it worked
please let me know. If not is there a product out there that is said to be
working in these conditions?
Gabriel
INRS-Institut Armand-Frappier
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 2
Date: Tue, 8 Jun 2004 11:10:01 -0600
From: "Patsy Ruegg" <pruegg <@t> colobio.com>
Subject: [Histonet] CORRECTIONLymphocytes antibodies - Fixed tissue
To: "Marceau, Gabriel \(IAF\)\(LAVAL\)"
<Gabriel.Marceau <@t> inrs-iaf.uquebec.ca>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <NGBBIMHHKLGNCOCPDKBOIELMCHAA.pruegg <@t> colobio.com>
Content-Type: text/plain; charset="iso-8859-1"
correction I meant to say that I use RABBIT monoclonal cd3 from Lab Vision.
Patsy
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Marceau,
Gabriel (IAF)(LAVAL)
Sent: Tuesday, June 08, 2004 8:53 AM
To: 'histonet <@t> lists.utsouthwestern.edu'
Subject: [Histonet] Lymphocytes antibodies - Fixed tissue
I Hi everyone,
I am looking for antibodies against lymphocytes (CD3 or other) that will
work on fixed tissue. Most of those comercially available are good for
cytometry but none is marked as usable on paraformaldehyde fixed tissue. If
someone tried one that is not specified for fixed tissue and it worked
please let me know. If not is there a product out there that is said to be
working in these conditions?
Gabriel
INRS-Institut Armand-Frappier
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 3
Date: Tue, 08 Jun 2004 19:28:17 +0200
From: Pablo S?nchez Quinteiro <psanquin <@t> lugo.usc.es>
Subject: RE: [Histonet] Double immunofluorescence with Alexa Fluor
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID: <3.0.6.32.20040608192817.007a1e90 <@t> pop.lugo.usc.es>
Content-Type: text/plain; charset="us-ascii"
Thanks for your kind answer Dave. I'll go for 633, but I am still worry.
Would not the donkey anti-goat IgG conjugate to the goat anti-rabbit IgG?
In my protocol both secondary antibodies are incubated together.
I am very grateful for any input.
Pablo
------------------------------
Message: 4
Date: Tue, 08 Jun 2004 13:23:56 -0400
From: Slydog5367 <@t> aol.com
Subject: [Histonet] JOB OPENING
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID: <59E05DCC.26A4A1C6.0CDBCF07 <@t> aol.com>
Content-Type: text/plain; charset=iso-8859-1
There is an immediate opening for a full time position in SE virginia, At
riverside Regional Medical center. We are a full service lab with A full
menu of immunos, ISH and FISH. Please repond to my personnel email or call
1-757-594-2885 and ask for Rod
thanks all
Rod slyter, HTL(ASCP), PA(AAPA)
Anatomical Pathology manager
------------------------------
Message: 5
Date: Tue, 8 Jun 2004 13:29:36 -0400
From: "Kevin Conway" <kconway2 <@t> uwo.ca>
Subject: [Histonet] re: Automated Image analysis
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <JOEJIOEIHODECIPHPIADIEJKCBAA.kconway2 <@t> uwo.ca>
Content-Type: text/plain; charset="US-ASCII"
Hi Yan,
I'm working on a macro series (SIMAR - Simple IMAging Routines) for Scion
Image (also a free download available at www.scioncorp.com).
The macro package I'm working on is available at:
https://sourceforge.net/projects/imageapps/
I'm using these macros for analysing a large number of images from
immunocytochemistry on muscle cells grown in culture, but should be
adaptable to most histochemical or immunohistochemical stains. I still have
to develop some documentation, but they are mostly self-explanatory, just
load a directory-worth of images into scion, then execute the macro.
Note that there is an immunofluorescence macro that inverts the image prior
to performing thresholding and area measurement. For most
immunocytochemistry I've done a decent threshold has been in the 100-150
range (contrary to the default of 40).
If you (or anyone) use them, I'd only ask to see the results and how
reliable the method is for you (I'm particularly interested in seeing the
error bars). If anyone is interested in contributing feel free to contact
me. The thesis is bogging me down a bit now but I'll try to get
documentation and a feature wish-list up soon.
Cheers,
Kevin
------------------------------
Message: 6
Date: Tue, 8 Jun 2004 12:32:29 -0500
From: "Kapoor, Sue" <Sue.Kapoor <@t> uhsi.org>
Subject: [Histonet] HPV results - probe vs. antibody
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<61E9F2400F53D5119CFC00508B44E33B019F55F9 <@t> khmcexch.uhsi.org>
Content-Type: text/plain; charset="iso-8859-1"
I have a formalin fixed, paraffin embedded block I sent out for HPV testing.
Can anyone explain these results to me:
HPV monoclonal antibody - positive
HPV polyclonal antibody - positive
HPV low risk probe - negative
HPV high risk probe - negative
The HPV monoclonal antibody used is Dako's, clone K1H8, I don't know what
the poly Ab. is.
I'm wondering how the Ab.'s can be positive but the more sensitive probes
are negative??????
lost.....
Sue Kapoor, HT (ASCP)
Histology Coordinator
Kenosha Medical Center
Kenosha, WI
262-653-5570
------------------------------
Message: 7
Date: Tue, 08 Jun 2004 13:40:06 -0500
From: "Drew Allan Roenneburg" <ROENN <@t> surgery.wisc.edu>
Subject: Re: [Histonet] Lymphocytes antibodies - Fixed tissue
To: <Gabriel.Marceau <@t> inrs-iaf.uquebec.ca>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <s0c5c1c5.008 <@t> gw.surgery.wisc.edu>
Content-Type: text/plain; charset=US-ASCII
Hi Gabriel
I have used Dakocytomation T-cell, CD3 (mouse monoclonal) on frozen
sections that were acetone fixed.
Drew Roenneburg
UW Medical School
Madison, WI
------------------------------
Message: 8
Date: Tue, 08 Jun 2004 15:26:44 -0400
From: DDittus787 <@t> aol.com
Subject: Re: [Histonet] HPV results - probe vs. antibody
To: Sue.Kapoor <@t> uhsi.org, histonet <@t> lists.utsouthwestern.edu
Message-ID: <63045660.5CBA70CA.0A1F969F <@t> aol.com>
Content-Type: text/plain; charset=iso-8859-1
Sue:
there are a variety of sub species (or viral groups that make up HPV-your
low risk and high risk are usually very specific ,while IHC antibodies are a
larger pool of types, hence, IHC positive ,probes negative.
not a complex answer but a basic one.
Dana
------------------------------
Message: 9
Date: Tue, 08 Jun 2004 15:27:14 -0400
From: DDittus787 <@t> aol.com
Subject: Re: [Histonet] HPV results - probe vs. antibody
To: Sue.Kapoor <@t> uhsi.org, histonet <@t> lists.utsouthwestern.edu
Message-ID: <72E5D6C3.6E5B0CF1.0A1F969F <@t> aol.com>
Content-Type: text/plain; charset=iso-8859-1
Sue:
there are a variety of sub species (or viral groups that make up HPV-your
low risk and high risk are usually very specific ,while IHC antibodies are a
larger pool of types, hence, IHC positive ,probes negative.
not a complex answer but a basic one.
Dana
------------------------------
Message: 10
Date: Tue, 8 Jun 2004 14:58:53 -0500
From: Julie Lee <labtech <@t> prairielakes.com>
Subject: [Histonet] IHC Staining Basics
To: "'histonet <@t> lists.utsouthwestern.edu'"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <490C2FC87D32D51196B100508BE3A05877D496 <@t> PYTHON>
Content-Type: text/plain; charset="iso-8859-1"
Hello from out here on the USA prairie!
We are a small hospital ( 1 full-time tech and 1 part-time tech). Our
pathologist would like us to start performing some more commonly requested
IHC stains (S-100, Vimentin, Cytokeratin, LCA). We are currently sending all
IHC to reference labs. This is a new undertaking for us. We are looking for
any suggestions as far as equipment (microwave, steamer ect.) and
methodologies for deparaffiniazation and antigen retrieval. The staining
will be done by hand. Any ideas for our situation would be greatly
appreciated!
Thanks,
JulieAnne
------------------------------
Message: 11
Date: Tue, 08 Jun 2004 21:41:05 +0000
From: "kevin williams" <akwilliams75 <@t> hotmail.com>
Subject: [Histonet] Reconditioned Leica Autostainer or Sikora Bach
Stainer
To: histonet <@t> pathology.swmed.edu
Message-ID: <BAY18-F73jjs2DfdN3M0008273a <@t> hotmail.com>
Content-Type: text/plain
Dear All:
I am looking to buy one of these, I have been to LabX and beyond but I
am still searching.
I have been told that the Leica Autostainer is for H&E, but I believe
it is also for immunos, am I thinking of different models? or am I
just wrong?
I have also been put on to Lamar Jones, Lamar if you are out there I
would love to talk to you, but do not seem able to get past his voice
mail, any help on the situation would be appreciated.
Can any of you give any references for either of these two options.
Thank you for your help.
Yours faithfully,
A. Kevin Williams
_________________________________________________________________
[1]Stop worrying about overloading your inbox - get MSN Hotmail Extra
Storage!
References
1. http://g.msn.com/8HMBENUS/2737??PS=47575
------------------------------
Message: 12
Date: Wed, 9 Jun 2004 08:04:17 +1000
From: "Rosalba" <zumbor <@t> email.cs.nsw.gov.au>
Subject: Re: [Histonet] Microscopic Bubbles........by the hundreds &
thousands
To: "Histonet" <histonet <@t> lists.utsouthwestern.edu>, "Tom Truscott"
<ttruscot <@t> vetmed.wsu.edu>
Message-ID: <003b01c44da4$8edc6400$1e7b4c98 <@t> HistoLab>
Content-Type: text/plain; charset="iso-8859-1"
We use Dako CD3 on fixed tissue
Original Message -----
From: "Tom Truscott" <ttruscot <@t> vetmed.wsu.edu>
To: "Bruce Abaloz" <brucea <@t> unimelb.edu.au>;
<histonet-bounces <@t> lists.utsouthwestern.edu>;
<histonet <@t> lists.utsouthwestern.edu>
Sent: Wednesday, June 09, 2004 12:53 AM
Subject: RE: [Histonet] Microscopic Bubbles........by the hundreds &
thousands
Bruce, Maybe you will have to fix the plant in formalin first to stop
all natural processes such as gas formation. Good luck, Tom T. USDA
Pullman, WA.
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Bruce
Abaloz
Sent: Monday, June 07, 2004 5:49 PM
To: histonet-bounces <@t> lists.utsouthwestern.edu;
histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Microscopic Bubbles........by the hundreds &
thousands
Dear Histonetters,
We have done the Oil Red O Charukian Method and Oil Red O technique
from Bancroft & Stevens, 3rd edition - both worked beautifully. Also
did Toluidine Blue stain combo with the Oil Red O & by itself - all
stunning results. Did the stains on cryo -sections of a native
Australian plant to demonstrate that the 'grainy' bits on the
underside of the leaves were full of Lipids/fats.
The PROBLEM we have encountered, is - no matter what Aqueous
Mountant we try , our cryosections are covered in microscopic air
bubbles (have tried to date: BDH/ Aquamount; DAKO /Faramount &
Glycergel & Ultramount.....even plain ol' Glycerol, (coverslipping
from DH20/Deionized H20/Tap H2O/Hot H2O/air dried - no H20).......
ALL resulted with same problem.
Can someone PLEASE advise what we are doing wrong (???) OR suggest a
possible answer as we need these results to be photographed ASAP.
Cheers & THANKS in advance,
Bruce in Melbourne/OZ
--
BRUCE ABALOZ PH:61383446282
HISTOLOGIST FAX:61383447909
DEPT.of ZOOLOGY EMAIL: brucea <@t> unimelb.edu.au
THE UNIVERSITY Of MELBOURNE.
VICTORIA.AUSTRALIA 3010
Nobody can make you feel inferior without your
permission. - Eleanor Roosevelt
DANCE LIKE NO-ONE'S WATCHING
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Message: 13
Date: Tue, 8 Jun 2004 15:23:13 -0700
From: "Lori Wilson" <WilsonL <@t> JWCI.ORG>
Subject: [Histonet] (no subject)
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <F24089984B27B048B962E34322A6FC3C070DFB <@t> smexsvr1.JWCI.ORG>
Content-Type: text/plain; charset="us-ascii"
We are staining parathyroid tissue as well as breast tissue for LCM. We
cut 4 - 6 um and plan to use it for protein extraction. Does anyone
have any experience using the histogene stain?
------------------------------
Message: 14
Date: Tue, 8 Jun 2004 17:40:08 -0700
From: "HCS" <int09018 <@t> alphahunt.com>
Subject: [Histonet] Thanks for the helping find antibody
To: "Histonet" <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <000701c44dba$514504b0$6501a8c0 <@t> hp>
Content-Type: text/plain; charset="iso-8859-1"
Thank you for your help, I have found several places to obtain this
information now.
>Hi, does anyone have a source for "murine CD11b and/or
TNF-alpha"?
>
>Also, the procedure for either murine CD11b and/or TNF-alpha?
LeRoy Brown HT(ASCP)HTL
HCS
www.histocs.com
1-360-966-7300
------------------------------
Message: 15
Date: Wed, 9 Jun 2004 08:21:13 +0100
From: "Garry Ashton" <GAshton <@t> PICR.man.ac.uk>
Subject: [Histonet] renal carcinoma cells
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<BAA35444B19AD940997ED02A6996AAE0176633 <@t> sanmail.picr.man.ac.uk>
Content-Type: text/plain; charset="us-ascii"
Dear all,
I have just been asked is there any way of identifying renal carcinoma
cells in tissue culture flasks.
The researcher is more interested confirming they are renal cells, and
not just carcinoma.
Any ideas?
Thanks in advance.
Garry
PICR
UK
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soon as possible.
------------------------------
Message: 16
Date: Wed, 09 Jun 2004 09:52:20 -0400
From: Jill Songer <jtsonger <@t> vt.edu>
Subject: [Histonet] charge for cryostat use
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <6.0.0.22.0.20040609094351.01af34b0 <@t> pop.vt.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed
Hi Histonetters,
We have a cryostat that lives in the histopath lab but we rarely use it.
However it is used by researchers both within the veterinary school and
other departments on the University campus. we have not charged for the use
of it, but are discussing instituting a charge to cover maintenance/repair
fees. My question is, is anybody out there experiencing the same? Do you
charge and if so, how much? Do you charge per hour or per use? The
researchers use their own slides and blades, so we don't need to charge for
those, but would like to recoup some of the money spent for recent repairs.
Thanks!!
Jill Songer HT (ASCP)
Histopathology Lab
Veterinary Medical Teaching Hospital
Virginia Tech
Blacksburg, VA 24061-0443
www.vetmed.vt.edu
------------------------------
Message: 17
Date: Wed, 9 Jun 2004 10:18:43 -0400 (EDT)
From: "SMITH,REBEKAH FELICIA" <rockbeki <@t> ufl.edu>
Subject: [Histonet] blocking endogenous peroxidase when using ABC
staining and DAB
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <1819142764.1086790723247.JavaMail.osg <@t> spnode37>
Content-Type: text/plain; format=flowed; charset=us-ascii
Hi,I'm working as a lab technician at UF making using
immunohistochemistry (ABC method and then DAB, specifically) to
find out where eNOS is located in sheep placenta tissue. However,
I seem to be both not getting staining on the endothelial cells
(which is weird since I'm using eNOS antibody) and getting a lot
of staining due to endogenous peroxidase activity. I've already
tried putting the slides in a solution of 1:10 30%hydrogen
peroxide to methanol for as long 40 min, and it still doesn't seem
to quench the endogenous peroxidase. Anyone have any suggestions
as to other methods of quenching it?
--
SMITH,REBEKAH FELICIA
"You are a child of the universe, no less than the trees and the
stars
You have a right to be here and whether or not it is clear to you,
no doubt the universe is unfolding as it should. Therefore be at
peace with G-d, whatever you conceive Him to be. And whatever your
labors and aspirations,in the noisy confusion of life, keep peace
in your soul.-Max Ehrmann,"Desiderata"
------------------------------
Message: 18
Date: Wed, 9 Jun 2004 10:59:05 -0400
From: " Katri Tuomala" <katri <@t> cogeco.ca>
Subject: Re: [Histonet] blocking endogenous peroxidase when using ABC
stainingand DAB
To: "SMITH,REBEKAH FELICIA" <rockbeki <@t> ufl.edu>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <000701c44e32$529d7a40$ce989618 <@t> hala4.on.cogeco.ca>
Content-Type: text/plain; charset="iso-8859-1"
Hi Rebekah,
It could be you are dealing with something else than endogenous peroxidase.
We need to know more detail of your method to try and think it through:
1.What animal is your primary antibody raised in and is it known to work on
sheep?
2.What are you using to block non-specific staining?
3.What animal is your secondary antibody raised in?
4. Are you working with formalin fixed, paraffin embedded or frozen
sections?
There are a lot of techs working on animal tissues on HistoNet. If you
provide more information, I'm sure help will be forthcoming...
Katri
Katri Tuomala
Department of Pathology
St.Joseph's Health Care
Hamilton,Ontario,Canada
----- Original Message -----
From: "SMITH,REBEKAH FELICIA" <rockbeki <@t> ufl.edu>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Wednesday, June 09, 2004 10:18 AM
Subject: [Histonet] blocking endogenous peroxidase when using ABC
stainingand DAB
> Hi,I'm working as a lab technician at UF making using
> immunohistochemistry (ABC method and then DAB, specifically) to
> find out where eNOS is located in sheep placenta tissue. However,
> I seem to be both not getting staining on the endothelial cells
> (which is weird since I'm using eNOS antibody) and getting a lot
> of staining due to endogenous peroxidase activity. I've already
> tried putting the slides in a solution of 1:10 30%hydrogen
> peroxide to methanol for as long 40 min, and it still doesn't seem
> to quench the endogenous peroxidase. Anyone have any suggestions
> as to other methods of quenching it?
> --
> SMITH,REBEKAH FELICIA
> "You are a child of the universe, no less than the trees and the
> stars
> You have a right to be here and whether or not it is clear to you,
> no doubt the universe is unfolding as it should. Therefore be at
> peace with G-d, whatever you conceive Him to be. And whatever your
> labors and aspirations,in the noisy confusion of life, keep peace
> in your soul.-Max Ehrmann,"Desiderata"
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 19
Date: Wed, 09 Jun 2004 08:01:57 -0700
From: Andrea Grantham <algranth <@t> u.arizona.edu>
Subject: Re: [Histonet] charge for cryostat use
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<4.3.2.7.2.20040609075725.00c6bd60 <@t> algranth.inbox.email.arizona.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed
I have pretty much the same scenario in my lab and a couple years ago we
instituted a $10/hr. charge. Last year cryostat use earned enough money to
buy a service contract for it.
Andi
At 09:52 AM 6/9/2004 -0400, you wrote:
>Hi Histonetters,
>
>We have a cryostat that lives in the histopath lab but we rarely use it.
>However it is used by researchers both within the veterinary school and
>other departments on the University campus. we have not charged for the
>use of it, but are discussing instituting a charge to cover
>maintenance/repair fees. My question is, is anybody out there experiencing
>the same? Do you charge and if so, how much? Do you charge per hour or per
>use? The researchers use their own slides and blades, so we don't need to
>charge for those, but would like to recoup some of the money spent for
>recent repairs.
>
>Thanks!!
>
>Jill Songer HT (ASCP)
>Histopathology Lab
>Veterinary Medical Teaching Hospital
>Virginia Tech
>Blacksburg, VA 24061-0443
>www.vetmed.vt.edu
>
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
.....................................................................
: 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 <@t> u.arizona.edu) :
:...................................................................:
http://www.cba.arizona.edu/histology-lab.html
------------------------------
Message: 20
Date: Wed, 9 Jun 2004 11:26:39 -0400
From: Latitia.E.Floyd <@t> gsk.com
Subject: [Histonet] Help
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <OFD4CDE0E9.B56BEAE2-ON85256EAE.00545EAF <@t> sb.com>
Content-Type: text/plain; charset=us-ascii
Hello Histonetters:
I am looking for a buffered formic acid decal solution that will work
fairly rapidly for femoral head and stifle joint bones from several
different species. I would be grateful to all who are willing to share
product name,where it can be obtained, time in solution and processing
times.
Much Appreciated,
Tish Floyd
Latitia .E. Floyd @ GSK.Com
GlaxoSmithKline
709 Swedeland Road
UEO461
Safety Assessment
King of Prussia, PA
19406
USA
Tel: 610-270-7629
Fax: 610-270-7202
Pager: 610-963-6688
------------------------------
Message: 21
Date: Wed, 09 Jun 2004 10:45:22 -0500
From: "Linda Jones" <ljones <@t> pathology.umsmed.edu>
Subject: [Histonet] Toe nails !!!!!
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <s0c6ea51.091 <@t> GWIA1.UMSMED.EDU>
Content-Type: text/plain; charset=US-ASCII
Hi,
Do anyone know how to section and stain toe nails? The nails are hard
to cut and the nail probably won't stay on the slide.
Thanks
Linda Harper-Jones,BS.HT. HTL(ASCP)
University of Mississippi Medical Center
Department of Pathology
Chief Histotechnologist Supervisor
(601) 984-1576
Linda Harper-Jones,BS.HT. HTL(ASCP)
University of Mississippi Medical Center
Department of Pathology
Chief Histotechnologist Supervisor
(601) 984-1576
------------------------------
Message: 22
Date: Wed, 9 Jun 2004 09:45:41 -0600
From: "Elizabeth Chlipala" <lizchlipala <@t> premierhistology.com>
Subject: RE: [Histonet] Help
To: <Latitia.E.Floyd <@t> gsk.com>, <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <000001c44e38$d51e3bc0$74d48a80 <@t> LIZ>
Content-Type: text/plain; charset="us-ascii"
Latitia
I make up my own formic acid, its much cheaper than commercially
available products. I will use concentrations from 5 to 20 percent
depending on how quick I need to get the projects done. I have some
images on my web page of mouse, rat and guinea pig joints. But I have
used formic acid on dog, horse and sheep joints also. If you have any
additional questions give me a call or e-mail me back.
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)
Premier Histology Laboratory, LLC
P.O. Box 18592
Boulder, Colorado 80308
Office: (303) 735-5001
Fax: (303) 735-3540
lizchlipala <@t> premierhistology.com
www.premierhistology.com
Ship to Address:
Premier Histology Laboratory
University of Colorado
MCBD, Room A3B40
Boulder, Colorado 80309
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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Latitia.E.Floyd <@t> gsk.com
Sent: Wednesday, June 09, 2004 8:27 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Help
Hello Histonetters:
I am looking for a buffered formic acid decal solution that will work
fairly rapidly for femoral head and stifle joint bones from several
different species. I would be grateful to all who are willing to share
product name,where it can be obtained, time in solution and processing
times.
Much Appreciated,
Tish Floyd
Latitia .E. Floyd @ GSK.Com
GlaxoSmithKline
709 Swedeland Road
UEO461
Safety Assessment
King of Prussia, PA
19406
USA
Tel: 610-270-7629
Fax: 610-270-7202
Pager: 610-963-6688
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 23
Date: Wed, 09 Jun 2004 11:56:31 -0400
From: Chris Bjornsson <cbjornss <@t> wadsworth.org>
Subject: [Histonet] Rat brain vascular casting using Mercox
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <BCECAB6F.B8D%cbjornss <@t> wadsworth.org>
Content-Type: text/plain; charset="US-ASCII"
Hi Histonetters,
I've been trying to get complete/satisfactory Mercox casts of rat cerebral
cortex, and have spent considerable amounts of time and reagents
troubleshooting with limited success. A difficult technique! We want to be
able to compare results among different animals and have therefore been
using a constant-pressure perfusion system. I've been performing
transcardiac perfusions with 37degC heparinized buffer, followed by
room-temp 4% PF fixation so we can stain different cell structures, and
using rhodamine B-labeled Mercox diluted 20% with MMA. I've tried changing
pressures, fixed vs. unfixed tissue, clamping the descending aorta, etc. to
no avail. So far it appears that Mercox pressure is actually less critical
than the flow rate of buffer & fix, and that clamping the descending aorta
improves vessel filling but doesn't completely solve our problems. We've
also noticed that little things like perfusion needle angle and placement
have a profound impact on flow rate and cast quality. I haven't explored
temperature effects much as of yet. Does anyone have any experience or
advice on this topic?
Thanks, cheers,
Chris Bjornsson
Laboratory of Nervous System Disorders
Wadsworth Center
Albany, NY 12201
------------------------------
Message: 24
Date: Wed, 09 Jun 2004 10:12:43 -0600
From: Gayle Callis <gcallis <@t> montana.edu>
Subject: Re: [Histonet] Help
To: Latitia.E.Floyd <@t> gsk.com, Histonet <@t> lists.utsouthwestern.edu
Message-ID: <3.0.6.32.20040609101243.00c1d560 <@t> gemini.msu.montana.edu>
Content-Type: text/plain; charset="us-ascii"
Latitia,
There is no concentrated buffered formic acid recipe. IF you calculate %
formic acid in commercial solutions or from a recipe, it is only ~4.5%.
Buffered formic acid is good if you want to do IHC, however, it is
published on effects of decalcification that concentrated acid decalcifies
rapidly, and that in itself protects antigen.
Make up your own formic acid decalcifier, it does NOT have to be buffered
as long as you have bone that is totally fixed in NBF. The main thing is
to do a decalcification endpoint check to insure bone is not overexposed to
your acid decalcifier.
You can use 10- 15% formic acid, made from 90% stock solution. 15 mls of
90% stock formic acid and qs to 100 mls with distilled water. Use at least
20 times volume of solution to size of bone.
Ways to speed up decalcification with 15% formic acid
Reduce size of bone, cut into slabs 5 mm thick, too thin is not good for
paraffin sectioning, bone will chip out of block. Do not stir if you do a
chemical endpoint test. You need to take aliquot off bottom of container.
Do not put two pieces of bone in same solution when doing this. Hopefully
you have xray for endpoint check, most sensitive method, excellent,
although chemical or a weight loss/weight gain method will work.
Suspend bone in a bag, cheesecloth in the decalcifier. Rinse bone next day
briefly to get calcium residing on surface of bone off and CHANGE to fresh
decalcifier to replenish formic acid.
At 11:26 AM 6/9/2004 -0400, you wrote:
>Hello Histonetters:
>
>I am looking for a buffered formic acid decal solution that will work
>fairly rapidly for femoral head and stifle joint bones from several
>different species. I would be grateful to all who are willing to share
>product name,where it can be obtained, time in solution and processing
>times.
>
>
>Much Appreciated,
>
>Tish Floyd
>
>
>Latitia .E. Floyd @ GSK.Com
>GlaxoSmithKline
>709 Swedeland Road
>UEO461
>Safety Assessment
>King of Prussia, PA
>19406
>USA
>Tel: 610-270-7629
>Fax: 610-270-7202
>Pager: 610-963-6688
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)
------------------------------
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet
End of Histonet Digest, Vol 7, Issue 13
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