RE: Daily Digest
Hello Jill,
Mounting medium jars can be found at www.tedpella.com
#21033 Mounting Medium/Balsam Bottle, ea.
#21033-6 Mounting Medium/Balsam Bottle, cs/6
Sharon Harvey
Naval Hospital
Charleston, SC 29405
sharvey@charleston.med.navy.mil
-----Original Message-----
From: HistoNet Server [mailto:histonet@pathology.swmed.edu]
Sent: Thursday, March 27, 2003 12:55 AM
To: HistoNet Server
Subject: Daily Digest
---------------------------------------------------------------------
Date: 26 Mar 2003 01:15:50 -0600
From: "J. A. Kiernan"
Subject: Counterstaining after in situ hybridization
Grrr!
First. You asked a question with a blank Subject line.
For this you deserve castration without anaesthesia etc.
That said, here's a possible answer to your question.
The RNase used to remove molecules of your unbound probe also removes
ribosomal RNA, which is what you were trying to stain with cresyl violet.
That's why you can't do a Nissl stain - the Nissl substance (rRNA) has been
removed.
In my lab about 10 years ago we did some in situ with 35S on rats' medullas
and encountered this problem, which we should have anticipated. The solution
was simple: stain the basic proteins that accompany nucleic acids and are
not attacked by nucleases. There was plenty of published guidance, with
Lillie's bigger book (either of its last 2 editions) carrying plenty of
references.
We did this (Paragraphs quoted from the M&M in the MS of a paper):
The slides were coated with Kodak NTB-2 nuclear track emulsion, and
autoradiographs were prepared. The exposure time was 22-days. Some
sections were counterstained with 0.1% fast green FCF at pH9.0 for 1h,
washed, dehydrated, cleared and mounted in DPX, a resinous medium. The
counterstain was to show the shapes of the cells by virtue of their basic
nucleoproteins [16], which had been unmasked by the RNase treatment.
[Ref 16]
16 James J, Tas J. Histochemical Protein Staining Methods (Royal
Microscopical Society Microscopy Handbooks 04). Oxford: Oxford
University Press, 1984: 28-29
We wrote this up, and your library should have the journal:
Savedia, S. & Kiernan, J. A. 1994. Increased synthesis of ubiquitin mRNA in
motor neurons after axotomy. Neuropathology and Applied Neurobiology 20,
577-586.
The paper has more than 50 references, many of them relating to your
question.
(Sheila Savedia was the MSc student who did most of the lab work. She went
on to qualify as a teacher and then got into a medical school. I don't know
where she is now.)
- --
- -------------------------
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/
_____________________________________________________________
sanjay rakhade wrote:
>
> Hi everyone,
>
> I am trying to set up a protocol for
> counterstaining emulsion dipped slides (for in situ
> hybridization using 35S labelled radioprobe) with cresyl violet
> stain. I tried to do the same using 1% cresyl violet acetate
> for 15 min followed by dehydration in serial 50, 70, 80, 95 and
> 100% Ethanol and dipping in Xylene followed by Permamount. The
> brain sections on my slide have the in situ grains intact,
> however the cresyl violet stains the cells, especially the
> neurons on the sections very lightly. I am not sure if this is
> due to the emulsion , dehydration or some problem with my
> staining protocol. I would appreciate it , if someone on this
> list has tried this experiment before and can help me
> troubleshoot ( or perhaps post a protocol for the same).
>
> Thanks in advance,
>
> Sanjay
__________________________________________________________________________
----------------------------------------------------------------------
Date: 26 Mar 2003 02:00:18 -0600
From: Chris van der Loos
Subject: RE: Rat BRDU question
Dear Nick,
Concerning your double staining problem on dog pancreas combining BrdU and
insulin IHC:
You didn't mention the use of FFPE sections, but I guess this is the case.
As I wrote here some time ago you may use just HEIR with citrate pH6.0,
instead of using the harsh treatment with HCl. Very likely HIER will not
affect your insulin or glucagon staining, and if you are lucky it is even
enhancing the staining.
Good luck!
Chris van der Loos
Dept.of Cardiovascular Pathology
Academic Medical Center
Amsterdam, The Netherlands
Nick Madary wrote:
>Date: 25 Mar 2003 15:30:27 -0600
>From: Nick_Madary@hgsi.com
>Subject: Rat BRDU question
>
>Pancreatic Islets on rat pup pancreas(7 day old pups), we can barely
>harvest pancreas anyway, and at that age an insulin shows up as little
>dots if we are lucky, forget an actual Islet. So the pancreas is a
>growing pancreas because it is from a 7 day old rat pup, the BRDU lights
>up all over. We want to count the BRDU positive cells in just the
>Islets(in this case anything that would be insulin positive). So I have
>tried doing the insulin first then brdu and vice versa using different
>colored DAB. It is my feeling that the harsh treatment of the HCL and to
>a lesser extent the sodium borate in the BRDU is destroying any chance of
>getting the insulins to light up. I tried doing some regular special for
>pancreatic islets but I still run in to the problem of no real islets at
>the 7 day mark for a rat.
>So how can I count BRDU cells in the areas I feel confident is the
>pancreatic islet only. Also did a glucagon/BRDU doblestain and it did not
>work either. I do not feel an Alk Phos/HRP is even the issue here.
----------------------------------------------------------------------
Date: 26 Mar 2003 02:45:23 -0600
From: sebres
Subject: Re: Counterstaining after in situ hybridization
What we do routinely in NTB2 emulsion-dipped brain sections after ISH with
35S is to use thionin as a counterstain after the slides are developed in
D19. An excellent protocol for this can be found at:
http://intramural.nimh.nih.gov/lcmr/snge/Protocols/ISHH/ISHH.html
Thionin is also a Nissl stain though, is it not? True, in a protocol for
ISH with an oligonucleotide probe, rather than a ribonucleotide probe, no
RNase would be involved. But even for ribo's, thionin works fine. The
stain is light, but this is desirable, as otherwise the stain would obscure
the silver grains. Am I missing something basic here? Susan Bachus - -----
Original Message -----
From: "J. A. Kiernan"
To: "sanjay rakhade"
Cc:
Sent: Wednesday, March 26, 2003 2:04 AM
Subject: Counterstaining after in situ hybridization
> Grrr!
> First. You asked a question with a blank Subject line.
> For this you deserve castration without anaesthesia etc.
> That said, here's a possible answer to your question.
>
> The RNase used to remove molecules of your unbound probe also
> removes ribosomal RNA, which is what you were trying to stain
> with cresyl violet. That's why you can't do a Nissl stain - the
> Nissl substance (rRNA) has been removed.
>
> In my lab about 10 years ago we did some in situ with 35S on
> rats' medullas and encountered this problem, which we should have
> anticipated. The solution was simple: stain the basic proteins
> that accompany nucleic acids and are not attacked by nucleases.
> There was plenty of published guidance, with Lillie's bigger book
> (either of its last 2 editions) carrying plenty of references.
>
> We did this (Paragraphs quoted from the M&M in the MS of a
> paper):
>
> The slides were coated with Kodak NTB-2 nuclear track emulsion,
> and autoradiographs were prepared. The exposure time was
> 22-days. Some sections were counterstained with 0.1% fast green
> FCF at pH9.0 for 1h, washed, dehydrated, cleared and mounted in
> DPX, a resinous medium. The counterstain was to show the shapes
> of the cells by virtue of their basic nucleoproteins [16], which
> had been unmasked by the RNase treatment.
>
> [Ref 16]
> 16 James J, Tas J. Histochemical Protein Staining Methods (Royal
> Microscopical Society Microscopy Handbooks 04). Oxford: Oxford
> University Press, 1984: 28-29
>
> We wrote this up, and your library should have the journal:
> Savedia, S. & Kiernan, J. A. 1994. Increased synthesis of
> ubiquitin mRNA in motor neurons after axotomy. Neuropathology and
> Applied Neurobiology 20, 577-586.
> The paper has more than 50 references, many of them relating to
> your question.
> (Sheila Savedia was the MSc student who did most of the lab work.
> She went on to qualify as a teacher and then got into a medical
> school. I don't know where she is now.)
> --
> -------------------------
> 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/
> _____________________________________________________________
>
> sanjay rakhade wrote:
> >
> > Hi everyone,
> >
> > I am trying to set up a protocol for
> > counterstaining emulsion dipped slides (for in situ
> > hybridization using 35S labelled radioprobe) with cresyl violet
> > stain. I tried to do the same using 1% cresyl violet acetate
> > for 15 min followed by dehydration in serial 50, 70, 80, 95 and
> > 100% Ethanol and dipping in Xylene followed by Permamount. The
> > brain sections on my slide have the in situ grains intact,
> > however the cresyl violet stains the cells, especially the
> > neurons on the sections very lightly. I am not sure if this is
> > due to the emulsion , dehydration or some problem with my
> > staining protocol. I would appreciate it , if someone on this
> > list has tried this experiment before and can help me
> > troubleshoot ( or perhaps post a protocol for the same).
> >
> > Thanks in advance,
> >
> > Sanjay
> __________________________________________________________________________
>
----------------------------------------------------------------------
Date: 26 Mar 2003 04:00:21 -0600
From: Fernando Capela e Silva
Subject: Mercury bromophenol blue method
I would like if someone send me the protocol of mercury bromophenol blue
method for proteins staining
With compliments
* * * * * * * * * * * * * * * * * * * * * * * * *
Fernando Capela e Silva
Laborat#243#rio de Biologia da Conserva#231##227#o
Departamento de Biologia
Universidade de ...vora
Apartado 94
7002-554 ...vora
PORTUGAL
Phone: +351-266 760 800
Fax: +351-266 711 231
Email: fcs@uevora.pt
http://evunix.uevora.pt/~fcs/FernandoCapelaSilva.htm
----------------------------------------------------------------------
Date: 26 Mar 2003 05:15:55 -0600
From: Myri37@aol.com
Subject: osteoblats
Hi everyone
Do you know any methods in staining or immunostaining, to selectively
display
osteoblats in decalcified tissue
thank you in advance
myriam
----------------------------------------------------------------------
Date: 26 Mar 2003 08:15:55 -0600
From: Janice A Mahoney
Subject: Re: CLIA Inspection
Hi Joe,
We had a CLIA inspection about a month after we had a Cap inspection at one
of our system hospitals too. We passed CAP with distinction. I think it is
becomming a common practice for CLIA to inspect labs after CAP.
There are a few differences in CAP and CLIA that I was unaware of. For
instance, CLIA defines how often Final/Frozen correlation is to be done ,
CAP doesn't state how often, just that it be performed. Good Luck with the
inspection.
Jan Mahoney
Omaha, NE
>>> Joe Nocito 03/25/2003 6:38:16 PM >>>
I guess this will teach me to have surgery and be out 3 weeks. I went to
work today to let them know that my surgeon released for work on next
Monday. I was told CLIA is coming next month.
Here's my question. Has anyone heard of CLIA performing an inspection two
months after CAP just on a routine basis? My CLIA experience is usually
someone squealed on the lab because of a disgruntled employee or to get
someone in trouble.
I was told that our CLIA license is up this year and that CLIA is trying to
come in after CAP to inspect. Why? We didn't receive any discrepancies on
CAP, why would CLIA want to inspect unless someone tipped them off.
I know this may be hard to believe, but some people in town hate my guts and
would love to see me squirm. Any one else experience this scenario?
Thanks.
Joe Nocito BS, HT (ASCP) QIHC
Histology Manager
Pathology Reference Lab
San Antonio, Texas
----------------------------------------------------------------------
Date: 26 Mar 2003 09:15:54 -0600
From: KMB1904@aol.com
Subject: staining issue
Hello histonetters!
We have been experiencing a problem in our lab with our staining...We
usually put 2 levels of bx speicmens side by side on a slide...we are having
problems with the level that ends up on the bottom in the staining rack.
This level stains lighter than the top level...our solutions are all above
the staining rack..so we dont feel that is the problem. Even when the
solutions are fresh this seems to occur..We are currently using Richard
allen products...Does anyone have a clue on this strange problem?
Thank you in advance!
Kathy Bowden
Nanticoke Memorial Hospital
Seaford De.
----------------------------------------------------------------------
Date: 26 Mar 2003 09:45:21 -0600
From: Vinnie Della Speranza
Subject: Re: staining issue
Kathy,
I have a hunch but need a bit more info.
you didn't mention if you stain the slides on a machine and if they are
stained with the slide in a vertical position? please indicate. also, are
you doing a regressive H&E, i.e. do you differntiate with acid alcohol??
Lastly, does this happen on all slides and have you ruled out the
possibility of a microtome problem, ie alternating thick and thin sections??
Vinnie
Vinnie Della Speranza
Manager for Anatomic Pathology Services
Medical University of South Carolina
165 Ashley Avenue Suite 309
Charleston, SC 29425
Ph: 843-792-6353
fax: 843-792-8974
>>> 03/26/03 09:55AM >>>
Hello histonetters!
We have been experiencing a problem in our lab with our staining...We
usually put 2 levels of bx speicmens side by side on a slide...we are having
problems with the level that ends up on the bottom in the staining rack.
This level stains lighter than the top level...our solutions are all above
the staining rack..so we dont feel that is the problem. Even when the
solutions are fresh this seems to occur..We are currently using Richard
allen products...Does anyone have a clue on this strange problem?
Thank you in advance!
Kathy Bowden
Nanticoke Memorial Hospital
Seaford De.
----------------------------------------------------------------------
Date: 26 Mar 2003 10:00:36 -0600
From: Tara Miller
Subject: Re: Fw: SAKURA COVERSLIPPER-experience
We've had our Sakura coverslipper for almost four years and we love it.
We've been told not to let the tissue dry out to prevent "corn flaking". So,
we don't put a second basket on to coverslip until the first is almost all
the way through. So far, So good.
Tara Oakes
>From: DeBari@mail.holyname.org
>To: Montse Verd#250# (Histopat)
>CC: Histonet
>Subject: Re: Fw: SAKURA COVERSLIPPER-experience
>Date: Mon, 24 Mar 2003 11:21:00 -0500
>
>works great, however if you load too many slides at once you get
>cornflaking. we've increased the # of xylene drips, but that doesn't seem
>to be the problem.
>christina debari
>holy name hospital
_________________________________________________________________
Protect your PC - get McAfee.com VirusScan Online
http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963
----------------------------------------------------------------------
Date: 26 Mar 2003 10:01:06 -0600
From: Ian Montgomery
Subject: Mould release.
After what seems like an eternity I'm finally running out of the proprietary
mould release I use. What's next, re-order or use homemade. As you'd expect
the bottle doesn't give much away, releasing agent in isopropyl alcohol.
Anybody using homemade and what is it? At the back of my mind I remember
people simply using a dilute washing up solution but is there something
better?
Ian.
Dr. Ian Montgomery,
Histotechnology,
Graham Kerr Building,
Institute of Biomedical & Life Sciences,
University of Glasgow,
Glasgow,
G12 8QQ.
Tel: 0141 339 8855
Office: 4652
Lab: 6644.
Pager: 07625 702883
e-mail: ian.montgomery@bio.gla.ac.uk
----------------------------------------------------------------------
Date: 26 Mar 2003 10:15:39 -0600
From: Gary Gill
Subject: RE: CLIA Inspection
CLIA '88 is Clinical Laboratory Improvement Amendments; CLIA '67 was
Clinical Laboratory Improvement Act. See:
http://www.phppo.cdc.gov/CLIA/chronol.asp
.
Gary Gill
- -----Original Message-----
From: Mike Konopka [mailto:mkonopka@scfa.edu.au]
Sent: Tuesday, March 25, 2003 8:34 PM
To: Histonet (E-mail)
Subject: RE: CLIA Inspection
Joe, sound like a lot of fun - not. Keep us posted as to the outcome.
Mike
- -----Original Message-----
From: Joe Nocito [mailto:jnocito@satx.rr.com]
Sent: Wednesday, 26 March 2003 2:15 PM
To: Mike Konopka
Subject: Re: CLIA Inspection
Mike,
CLIA is Clinical Laboratory Improvement Act implemented by the US government
in 1988. This is the government agency that can shut down a lab in a heart
beat. Although I have every thing ready to go, CLIA usually doesn't inspect
unless there is a complaint.
See, two inspectors cover all the labs in south Texas, which covers a vast
amount of territory and many hospital and private labs. And this is the same
inspector that inspected me at my other hospital a couple of years ago.
He's a real sweetheart. I hope he doesn't remember me. We had a little
disagreement the last time we met.
Joe
- ----- Original Message -----
From: Mike Konopka
To: Histonet (E-mail)
Sent: Tuesday, March 25, 2003 3:07 PM
Subject: RE: CLIA Inspection
Joe, sounds like there is a little bit of paranoia going on, view the CLIA
inspection as a positive. "unless someone tipped them off" is a worry - is
there something to hide. PS I don't know what CLIA stands for.
- -----Original Message-----
From: Joe Nocito [mailto:jnocito@satx.rr.com]
Sent: Wednesday, 26 March 2003 11:38 AM
To: Histonet
Subject: CLIA Inspection
I guess this will teach me to have surgery and be out 3 weeks. I went to
work today to let them know that my surgeon released for work on next
Monday. I was told CLIA is coming next month.
Here's my question. Has anyone heard of CLIA performing an inspection two
months after CAP just on a routine basis? My CLIA experience is usually
someone squealed on the lab because of a disgruntled employee or to get
someone in trouble.
I was told that our CLIA license is up this year and that CLIA is trying to
come in after CAP to inspect. Why? We didn't receive any discrepancies on
CAP, why would CLIA want to inspect unless someone tipped them off.
I know this may be hard to believe, but some people in town hate my guts and
would love to see me squirm. Any one else experience this scenario?
Thanks.
Joe Nocito BS, HT (ASCP) QIHC
Histology Manager
Pathology Reference Lab
San Antonio, Texas
---------------------------------------------------------------------
Date: 26 Mar 2003 10:30:46 -0600
From: "Grant, Debra"
Subject: VT1000
Hello Histonet,
We want to know if this model can cut the fresh live tissue for
later tissue culture. Thanks!
Debby Grant
Research Technician II
Histology Core Facility
Stowers Institute for Medical Research
1000 E. 50th Street
Kansas City, MO 64110
drg@stowers-institute.org
----------------------------------------------------------------------
Date: 26 Mar 2003 10:34:39 -0600
From: Gary Gill
Subject: RE: CLIA Inspection
Think of a post-CAP CLIA follow-up inspection as CLIA's quality assurance
method that CAP is doing its job as a deemed authority. Think positive; no
reason to be paranoid (i.e., a pair of noids). Besides, well run labs
should pass any inspection by any organization at any time, even unannounced
inspections.
Gary Gill
----------------------------------------------------------------------
Date: 26 Mar 2003 11:15:57 -0600
From: Terri Thinnes
Subject: staining too weak for ISH
Message for Sanjay about weak counterstaining of is situ hybridization
slides
Hi, I do a H&E counterstain on my radio-labelled in situ slides. But I
found the hematoxylin was too weak so I purchased a Gill triple strength
and an intensified eosin. I stain for 1 minute each. The stain is
beautiful. In comparison, if I use this same hematoxylin for an IHC
counterstain, I can do one quick dip and that is more than enough! Staining
5-10 minutes in Mayer's Hematoxylin did not stain intensely enough for my
ISH. So, I think you need a more concentrated stain. Terri Thinnes The
Scripps Research Institute
----------------------------------------------------------------------
Date: 26 Mar 2003 11:16:24 -0600
From: Laurel Baccei
Subject: Histotech Job Opportunity
I realize this is my second request, but I was hoping I would get a better
response the second time around. I apologize for the repetition.
I am searching for a certified Histotech (HT) for a full time employment
opportunity at a Contract Research Organization in Indianapolis, Indiana.
The company is looking for someone with 5-10 years experience in Histology.
The key requirement is extensive experience with high volumes of animal
tissue sampling. Degrees are not as important as the certification and
experience.
The company is offering relocation and a very competitive salary. Please
forward this to anyone who may be interested and feel free to contact me
with any questions or concerns.
Thanks for any and all leads in advance,
Laurel Baccei
SciTech Recruiters
650-323-3333 or 858-202-1700 Phone
laurel@scitechrecruiters.com
www.scitechrecruiters.com
"Leaders in the Recruitment of Scientific Professionals"
Date: 26 Mar 2003 11:46:04 -0600
From: Carol Bobrowitz
Subject: ventilation requirements
We are moving to a new lab.
I was wondering if OSHA, CAP or CLIA have the exact specifications for air
flow and ventilation requirements needed for a Histology Lab. If they do,
could anyone guide me to the correct source to obtain this information. (web
site or phone number would be helpful also)
Thank you for any information you could provide me.
Carol Ann Bobrowitz
Histology Laboratory
Department of Physiology
Medical College of Wisconsin
Milwaukee, Wisconsin
(414) 456-8179
FAX (414) 456-6546
cbobrowi@mcw.edu
Date: 26 Mar 2003 11:46:27 -0600
From: Sharon McLaughlin
Subject: Part-time work
Hi,
I failed to mention where I live. I live in the Seattle, Washington area
and am looking for evening and week-end work. I would like to work about 10
hours extra a week.
Thanks for your help,
Sharon McLaughlin
e
----------------------------------------------------------------------
Date: 26 Mar 2003 11:53:57 -0600
From: Pam Barker (extension 234)
Subject: Histo Tech needed in Big Sky Country
Hello Histonetters, We are working with an organization in Wyoming who is
in need of a histo tech. HT or HTL certified or eligible.
They offer a competitive salary, excellent benefits, relocation assistance
and a sign on bonus.
If you are interested in hearing more please contact me via e-mail at
pam@ategra.com or call me at 800-466-9919 x234..
Thank You !
Pam Barker, Sr. Recruiter
- --------------------------------------------
Ategra Systems Inc
Specialists in Permanent & Contract Staffing for the Clinical Lab
Professions
VOICE: 407-671-5800 ext 234
TOLLFREE: 800-466-9919 ext 234
FAX: 407-671-6075
EMAIL: pam@ategra.com
WEBSITE: www.ategra.com
- --------------------------------------------
----------------------------------------------------------------------
Date: 26 Mar 2003 12:30:33 -0600
From: edison narvaez
Subject: Tyrosinase Protocol
would anybody share their protocol for Tyrosinase, we have not been able to
get this AB to work.
We use the Dako Autostainer, water bath method for HIER, and envision
polymer from Dako.
Thank you....
- ---------------------------------
Do you Yahoo!?
Yahoo! Platinum - Watch CBS' NCAA March Madness, live on your desktop!
----------------------------------------------------------------------
Date: 26 Mar 2003 13:45:37 -0600
From: mari.ann.mailhiot@leica-microsystems.com
Subject: Re: VT1000
Debra
The purpose of the VT 1000S microtome is that it can be used to cut fresh
unfixed tissue. If you have any questions feel free to give me a call.
Regards
Mari Ann Mailhiot BA HT ASCP
Application Specialist
Leica Technical Assistance Center
800 248 0123 x7267
847 236 3063 fax
mari.ann.mailhiot@leica-microsystems.com
www.leica-microsystems.com
"Grant, Debra"
itute.org> cc:
Subject: VT1000
03/26/2003 10:16
AM
Hello Histonet,
We want to know if this model can cut the fresh live tissue for
later tissue culture. Thanks!
Debby Grant
Research Technician II
Histology Core Facility
Stowers Institute for Medical Research
1000 E. 50th Street
Kansas City, MO 64110
drg@stowers-institute.org
----------------------------------------------------------------------
Date: 26 Mar 2003 14:00:29 -0600
From: brett_connolly@merck.com
Subject: thanks for decloaker advice
Thanks to all for your input!!
Brett
Brett M. Connolly, Ph.D.
Merck & Co.,Inc.
MRL, Imaging
WP26A-3000
PO Box 4
West Point, PA 19486
PH 215-652-2501
fax. 215-652-2075
e-mail. brett_connolly@merck.com
-
----------------------------------------------------------------------------
--
Notice: This e-mail message, together with any attachments, contains
information of Merck & Co., Inc. (Whitehouse Station, New Jersey, USA) that
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message. If you are not the intended recipient, and have received this
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----------------------------------------------------------------------
Date: 26 Mar 2003 17:00:35 -0600
From: "Michelle D. Moore"
Subject: Hemolysis Standards
I know this is a histology forum but we have many diverse educational
backgrounds and I am trying to tap into those histotechs/medtechs brains. I
am asking this on behalf of my lab manager. She is trying to set up a policy
for specimen rejection by using a hemolysis standard. In Oct 1995 MLO put
out an article about Hemolysis meeting QIP and talked about a solution
called Drubbins solution.
What she was hoping for is finding a hemolysis standard that they can either
make or buy cheaply to stabilize the deterioration of cells. I hope this
covers what you need to know to help us out or direct me to someone or
someplace that can help us. Thank you in advance for you time and help.
Have a good today and a better tomorrow.
Michelle Moore HT(ASCP)
TMH
Craig, CO
----------------------------------------------------------------------
Date: 26 Mar 2003 17:30:10 -0600
From: pruegg@colobio.com
Subject: RE: Collagen staining
Terri,
I have a collagen staining procedure which does not use picric, it uses
Azure B.
Patsy
- -----Original Message-----
From: Terri Herbst [mailto:terriherbst@yahoo.com]
Sent: Tuesday, March 25, 2003 9:41 AM
To: histonet@pathology.swmed.edu
Subject: Collagen staining
I was wondering if anyone on this list had any
protocols or advice on a method of collagen staining
which doesn't involve picric acid. The institution I
work for is extremely against my lab using this
chemical. Many of the protocols I have come across
either include saturated picric acid solution or
Bouin's solution. Any suggestions?
Thank you,
Terri Herbst
Hines VA Hospital
Hines, IL
__________________________________________________
Do you Yahoo!?
Yahoo! Platinum - Watch CBS' NCAA March Madness, live on your desktop!
http://platinum.yahoo.com
----------------------------------------------------------------------
Date: 26 Mar 2003 17:45:09 -0600
From: "Morken, Tim"
Subject: Position opening at CDC Atlanta in near future
Histonetters,
Position opening at CDC in Atlanta
This is not an official announcement, but a friendly "heads-up". The remarks
here are completely unofficial and are only my personal observations.
Within the next few months, there will be a position opening for an
experienced Histotechnologist at the Centers for Disease Control in Atlanta,
GA, a US Federal Government agency under the Health and Human Services
Department. From what I understand at this point, the plan is to announce
the position as a Biologist and as a Medical Technologist. Pay grade is at
the GS-9 or GS-11 level, depending on education and experience. Please do
not let the position announcement titles fool you - the work is indeed more
related to a Histotechnologist position than a Med Tech. In order to apply
for the postion, any interested person will need to check the announcements
on the CDC job announcement website. Announcements are posted on Wednesdays
and Fridays. I will try to let you know when it is announced, but please
watch the job board too:
http://www.cdc.gov/hrmo/hrmo.htm
The ideal candidate will have a very strong background in routine and
specialized histology procedures. Immunohistochemistry expericence is not
necessary, but would be advantagous. Typically, a BA or BS degree in a
science field plus experience in histotechnology will be a minimum
requirement. HTL certification desirable, but not required. US citizenship
is required. The position will be in the histology section of the Infectious
Disease Pathology Activity (IDPA), which deals with all diagnostic tissue
samples sent to CDC and with all research projects involving tissues. In
addition to routine diagnostic activity, the group performs infectious
disease research and supports investigations of many infectious disease
outbreaks around the world. Recent examples of IDPA outbreak involvement
include Ebola, West Nile, anthrax, and SARS. Everyone in IDPA has the
opportunity to participate at many different levels of research and to learn
many different types of lab methods. Occasionally, laboratory staff are
offered the opportunity to present their work at national meetings and
attend training courses to enhance their skills. The only limitation is
that which the individual sets on themselves.
Staff at present: 4 pathologists, 5 histotechnolgists, 3 electron
microscopists, 2 molecular biologists, support staff, 2 research fellows,
several students.
Equipment in the lab: microm and leica microtomes, sakura DRS and DRS2000
H&E stainers, 3 DAKO and 1 Biogenex IHC stainers, 2 each TBS slide and
cassette labelers, 2 Sakura tissue processors, Hacker milestone microwave
tissue processor, 2 Shandon grosslab-senior grossing hoods, fully equipped
molecular biology lab, Zeiss laser capture microdissection instrument, DNA
sequencer, fully equipped electron microscopy suite with 3 TEMs. Each staff
member also has their own desk and computer.
I can say from personal experience that this is an excellent place to work.
The work is always interesting and challenging and new things come up every
week. There are no routine days. The group is very outgoing, and friendly
teamwork is the norm, although the individual can grow professionally as
much as they desire.
If you are in the Atlanta area, or could relocate, please consider this very
desirable position.
Tim Morken
CDC, Atlanta
----------------------------------------------------------------------
Date: 26 Mar 2003 17:45:34 -0600
From: pruegg@colobio.com
Subject: RE: osteoblats
Myriam,
Osteocalcin from Biomedical Technologies can be used for IHC labeling
osteoblasts in formic acid decalcified tissue.
Patsy
- -----Original Message-----
From: Myri37@aol.com [mailto:Myri37@aol.com]
Sent: Wednesday, March 26, 2003 4:03 AM
To: histonet@pathology.swmed.edu
Subject: osteoblats
Hi everyone
Do you know any methods in staining or immunostaining, to selectively
display osteoblats in decalcified tissue
thank you in advance
myriam
---------------------------------------------------------------------
Date: 26 Mar 2003 17:46:00 -0600
From: David Walker
Subject: Re: VT1000
On 3/26/03 9:16, "Grant, Debra" wrote:
>
> Hello Histonet,
>
> We want to know if this model can cut the fresh live tissue for
>
> later tissue culture. Thanks!
>
> Debby Grant
> Research Technician II
> Histology Core Facility
> Stowers Institute for Medical Research
> 1000 E. 50th Street
> Kansas City, MO 64110
> drg@stowers-institute.org
>
>
A researcher in our facility routinely uses a Leica VT1000S to cut mouse
brain sections for 3section cultures2.
David Walker
School of Pharmacy and Allied Health Sciences
University of Montana
Date: 26 Mar 2003 17:46:26 -0600
From: "Breckenridge, Richard A."
Subject: RE: Tyrosinase Protocol
We use clone T311 from NovoCastra. We use it at a dilution of 1:25. We also
use HIER for 20 min. in a pH 6.0 citrate buffer at 99 degrees and let cool
for 20 min. on the counter. We run it on the DAKO using Alk Phos. (we use
the Signet USA kit with Fast Red) We currently do not have a pic of this
stain on our website, however, we do have a lot of other pics and have added
nearly all the spec sheets (in .pdf format) for our antibodies. see them at
www2.utmb.edu/pps . Follow the "SPECIAL PROCEDURES" link on the left hand
side.
Sincerely,
Richard A. Breckenridge HT(ASCP)
Technical Supervisor
UTMB Special Procedures Laboratory
Clinical Sciences Building Room 504
Galveston, TX 77555-0748
Phone 409-772-8051 Fax 409-772-8030
- -----Original Message-----
From: edison narvaez [mailto:connhist@yahoo.com]
Sent: Wednesday, March 26, 2003 11:42 AM
To: histonet@pathology.swmed.edu
Subject: Tyrosinase Protocol
would anybody share their protocol for Tyrosinase, we have not been able to
get this AB to work.
We use the Dako Autostainer, water bath method for HIER, and envision
polymer from Dako.
Thank you....
Date: 26 Mar 2003 18:16:36 -0600
From: Barbara Stancel
Subject: Re: Mould release.
Dear Ian, We still prefer the purchased mold release, usually the stuff in
a pump. But in a pinch, I tried using spray vegetable oil (PAM brand). It
worked great! But we had to wash the grease off our blocks for a
while...there was just enough grease on the blocks to keep them from making
nice ribbons. We have also tried boiling the molds to clean them and using
them without any release spray. It also worked well, but we had to be sure
the blocks were very cold before we tried to get them out of the molds. Like
I said, we still prefer soaking them in xylene, dipping in acetone, drying
and spraying with purchased mold release.
Best of Luck.
Histologically yours,
Barbara H. Stancel, HTL(ASCP)HT
USDA, FSIS, OPHS, Eastern Laboratory, Pathology
RRC, 950 College Station Road
Athens, Georgia 30604
phone: (706) 546-3556
fax: (706) 546-3589
>From: Ian Montgomery
>To: histonet@pathology.swmed.edu
>Subject: Mould release.
>Date: Wed, 26 Mar 2003 15:48:00 +0000
>
> After what seems like an eternity I'm finally running out of the
>proprietary mould release I use. What's next, re-order or use homemade. As
>you'd expect the bottle doesn't give much away, releasing agent in
>isopropyl alcohol. Anybody using homemade and what is it? At the back of my
>mind I remember people simply using a dilute washing up solution but is
>there something better?
>Ian.
>
>Dr. Ian Montgomery,
>Histotechnology,
>Graham Kerr Building,
>Institute of Biomedical & Life Sciences,
>University of Glasgow,
>Glasgow,
>G12 8QQ.
>Tel: 0141 339 8855
>Office: 4652
>Lab: 6644.
>Pager: 07625 702883
>e-mail: ian.montgomery@bio.gla.ac.uk
_________________________________________________________________
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----------------------------------------------------------------------
Date: 26 Mar 2003 18:31:11 -0600
From: jill cox
Subject: Mounting medium jars
Hello everyone,
I have been looking for the mounting medium jars
for coverslipping and cant find anything except one
brand where you have to order a case at a time. They
are too expensive that way. Does anyone know where I
can get 2 to 3 of them? Thanks in advance
=====
Jill Cox HT (ASCP)
Seattle Histology Lab
Hello Jill,
Mounting medium jars can be found at www.tedpella.com
#21033 Mounting Medium/Balsam Bottle, ea.
#21033-6 Mounting Medium/Balsam Bottle, cs/6
Sharon Harvey
Naval Hospital
Charleston, SC 29405
sharvey@charleston.med.navy.mil
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