RE: Daily Digest
| From: | Debra Ritter <DRitter@stmarygj.com> |
Janet Maas works out of Fort Collins, Colo.
Sorry, thats all we know
Deb adn Bill in GJ, Co.
> ----------
> From: HistoNet Server[SMTP:histonet@pathology.swmed.edu]
> Sent: Wednesday, May 02, 2001 11:04 PM
> To: HistoNet Server
> Subject: Daily Digest
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> ----------------------------------------------------------------------
>
> Date: 2 May 2001 04:16:10 -0500
> From: "Neuropathology" <Neuropath.Frenchay@dial.pipex.com>
> Subject: Re: detergents in antibody diuents
>
> We have used Tween 20 in our antibody diluent at a concentration of
> 0.05%
> without any problems on both paraffin and frozen sections.
>
> Bob Quilty
> Dept of Neuropathology
> Frenchay Hospital
> Bristol UK
>
> - ----- Original Message -----
> From: Ron Salisbury <ronald2@uakron.edu>
> To: Histonet <histonet@pathology.swmed.edu>
> Sent: Tuesday, May 01, 2001 7:06 PM
> Subject: detergents in antibody diuents
>
>
> > I have seen various concentrations of detergents (Triton X, Tween 20
> > etc.) used in primary antibody diluents and wondered if there were any
> > guidelines for choosing a particular concentration. Does it depend on
> > whether paraffin or frozen sections are used or is it more dependent
> > upon the type and concentration of fixative? Also, is there an effect of
> > detergent concentration on the ability of sections to adhere to slides?
> >
> >
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 04:16:27 -0500
> From: "Neuropathology" <Neuropath.Frenchay@dial.pipex.com>
> Subject: Re: Immunostainer
>
> We also demoed (right spelling?) the Dako Horizon and the LabVision
> machines. Didn't like the Horizon, fiddly and required special slides etc.
> The LabVision was neat and clever with its Windows based software but we
> found patchy staining was a problem. It didn't seem to always cover the
> sections in spite of using detergent in the reagents.
>
> Bob Quilty
>
> - ----- Original Message -----
> From: David Addington-Hall <david.addington-hall@carmarthen.wales.nhs.uk>
> To: <Neuropath.Frenchay@dial.pipex.com>
> Sent: Tuesday, May 01, 2001 10:52 AM
> Subject: Re: Immunostainer
>
>
> I was interested in your email to Histonet about immunostainers.
> We are about to purchase an immunostainer, and seem to have a similar
> workload for immunos as you. I am aware of the Dako and Menarini machines,
> but would be grateful for information on any other suppliers.
>
> Thanks
>
> David Addington-Hall
> Lead BMS Histology/Cytology
> West Wales General hospital
> Carmarthen
> West Wales
>
> >>> Neuropathology <Neuropath.Frenchay@dial.pipex.com> 05/01 10:06 am >>>
> We demoed a number of immunostainers in 1999 and decided on the Optimax as
> the best of the bunch. We took delivery of it November 2000. We don't do a
> great number of immunos, varies from 30-60 a week, and the machine only
> dropped a pipette once. It is otherwise very reliable and once set up,
> easy
> to program runs. Works a treat with Biogenex liquid DAB which is stable
> for
> up to 8 hours.
>
> Bob Quilty
> Dept of Neuropathology
> Frenchay Hospital
> Bristol, UK.
>
> - ----- Original Message -----
> From: Brennan, Liam <Liam.Brennan@bll.n-i.nhs.uk>
> To: HISTONET <histonet@pathology.swmed.edu>
> Sent: Monday, April 30, 2001 12:01 PM
> Subject: Immunostainer
>
>
> > 30/04/01
> >
> > Anyone out there with an opinion/comment to make about either the
> Optimax
> or
> > the Labvision immunostainer's as both companies have tendered to supply
> us
> > with an immunostainer as well Ventana.
> >
> > many thanks
> > Liam Brennan
> > Histopathology Dept.
> > Belfast City Hospital
> >
> >
>
>
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 06:43:35 -0500
> From: Debra Lloyd <dlloyd@neuro.mcg.edu>
> Subject: Immunohistochemical staining on frozen muscle
>
> Hi everyone,
> I'm relatively new to the world of histology. I'm the Neuromuscular
> technician at the Medical College of GA in Augusta. My question pertains
> to Immunostaining of muscle biopsies on patients that may have muscular
> dystrophy. Therefore, I'm only doing dystrophin, sarcoglycan, merosin,
> and occasionally dysferlin. The problem that I'm experiencing is too
> much background staining and in the case of the gamma-sarcoglycan and
> dysferlin they're not working at all.
> I generally use the dystrophin kit supplied by Vector.
> Since the tissue thats used is small in quanity, I have been doing the
> procedure manually.
> The procedure used is:
> primary antibody (l hour) dilutions 1:300.
> rinse in TBS buffer (pH7.6) 3 x 10 minutes
> secondary antibody ( 1 hour) dilution 1:100.
> rinse in TBS buffer (pH 7.6) 3 x 10 minutes.
> DAB ( 4 minutes)
> counterstain harris hematoxylin.
> This is the protocol that Vector sends with its kit.
> Please let me know of any techniques, etc that will reduce the background
> staining.
> Thanks,
>
> Debra L. Lloyd, BS
> Neuromuscular Pathology Lab
> Medical College of GA
> phone: (706) 721-2312
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 07:02:12 -0500
> From: "Morken, Tim" <tim9@cdc.gov>
> Subject: RE: Re immunostainer
>
> We avoid this problem by ordering the demo unit through a no-cost purchase
> order which specifically states that it is being brought in for
> demonstration purposes only and we are under no obligation to buy the
> machine being demonstrated.
>
> Tim Morken, BA, EMT(MSA), HTL(ASCP)
> Infectious Disease Pathology Activity
> Centers for Disease Control and Prevention
> Ms-G32
> 1600 Clifton Road
> Atlanta, GA 30333
> USA
>
> PH: 404-639-3964
> FAX: 404-639-3043
>
> email: tim9@cdc.gov
>
>
>
> - -----Original Message-----
> From: Cynthia Tily [mailto:cynthiatily@mindspring.com]
> Sent: Tuesday, May 01, 2001 7:08 PM
> To: Clarke Ian; histonet
> Subject: Re: Re immunostainer
>
>
> Funny, we have this problem woth Ventana!
> - ----- Original Message -----
> From: Clarke Ian <clarke.ian@virgin.net>
> To: histonet <histonet@pathology.swmed.edu>
> Sent: Tuesday, May 01, 2001 4:50 PM
> Subject: Re immunostainer
>
>
> > Liam,
> > Be careful with the tender from the Optimax,They put a tender in with us
> > along with Ventana ,so we had to try the Optimax against the Ventana as
> is
> > required with the tendering process.However , Optimax had contacted our
> > purchasing department and asked for the money up front for a trial
> ,unknown
> > to us, of #163#20,000 .
> >
> > During the trial we found that we got uneven staining and despite
> several
> > attempts by Optimax we could not get the consistency/intensity of
> staining
> > that the ventana had.We then found out that the purchasing department
> had
> > paid the money out .Optimax were contacted to ask for the money back and
> we
> > have been in a battle ever since.We have the stainer in a store room
> > gathering dust.You can come up and see it if you wish
> >
> > Ian Clarke
> > Craigavon Area Hospital
> >
> >
> >
> >
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 07:26:57 -0500
> From: dl.botsford@sympatico.ca
> Subject: Re: PAP pens
>
> Hi Christine,
>
> Before we automated we used to circle the section with petroleum jelly (
> vaseline) to create the reagent well.
>
> Run the slides down to water. Fill a 10cc syringe with jelly. Carefully
> wipe
> around the section area to make it dry. Apply the jelly around the tissue
> with
> the syringe. Take a wet tissue paper and press the outside edge of the
> jelly
> against the slide to form the reagent well.This take a little practice.
> When
> finished the staining, run the slides in hot water and the jelly floats
> off.
> Counter stain with Heam. and coverslip as ususal.This is one of the
> "pearls of
> wisdom" that I picked up from a Hamilton , Ontario , Canada researcher in
> a
> hallway discussion at the NSH convention a few years back. They used very
> high
> dilution of a their homemade antibody to conserve on the antibody and
> incubated in the frig for 2 days. By placing a coverslip on the vaseline
> she
> created a humidity chamber which allowed the section to remain moist
> during
> the incubation.
>
> Sincerely,
> Dan Botsford
> Windsor Regional Hospital
> Windsor, Ontario, Canada
>
> cmcbride@tulane.edu wrote:
>
> > Hi,
> > Does anyone have a better alternative to PAP pens? Their membrane seems
> to
> > breakdown after a few soln changes.
> > Thanks,
> > Christine
> >
> > Christine McBride, M.S.
> > Tulane University Medical Center
> > Center for Gene Therapy
> > SL-99
> > 1430 Tulane Ave.
> > New Orleans, LA 70112
> > cmcbride@tulane.edu
> > Phone:(504) 988-7069
> > Fax:(504) 588-5326
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 07:27:19 -0500
> From: Robert Geske <RGeske@lexgen.com>
> Subject: ribo probes
>
> Does anyone have a commercial source for dig or FITC labeled riboprobes? i
> would need both sense and anti-sense. i have sources for oligos, but i
> specifically want labeled sense and anti-sense RNA.
>
> thanks in advance,
> rob****** The contents of this communication are intended only for the
> addressee and may contain confidential and/or privileged material. If you
> are not the intended recipient, please do not read, copy, use or disclose
> this communication and notify the sender. Opinions, conclusions and other
> information in this communication that do not relate to the official
> business of my company shall be understood as neither given nor endorsed
> by
> it. *****
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 08:07:57 -0500
> From: Robert Geske <RGeske@lexgen.com>
> Subject: histology services
>
> Does anyone know how to contact the histology service in Colorado that I
> believe is named Maas Histology. I would appreciate the phone number.
>
>
> thanks
>
> rob
>
> ****** The contents of this communication are intended only for the
> addressee and may contain confidential and/or privileged material. If you
> are not the intended recipient, please do not read, copy, use or disclose
> this communication and notify the sender. Opinions, conclusions and other
> information in this communication that do not relate to the official
> business of my company shall be understood as neither given nor endorsed
> by
> it. *****
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 08:08:13 -0500
> From: Roger Moretz <stamptrain@yahoo.com>
> Subject: Re: Mouse Perfusions
>
> Richard:
> Mouse perfusions are not terribly different, except
> maybe for the necessity of using finer needles. We
> use a saline drip bottle at about a 3 ft height above
> the table. Actually, there are 2 bottles--one filled
> with saline, and the second with the fixative. Fed
> into a common line at a 'T' or 'Y', it is possible
> then to easily switch from the saline flush to the
> fixative. The canula is either fine teflon tubing or
> a blunt needle 21 ga or smaller (I have used 30 ga on
> very young animals). The canula can be inserted into
> the left ventricle of the heart or into the abdominal
> aorta, and cut a vein for exit. You can use a bit
> higher level for the drip bottles (I am drawing a
> mental blank on the optimum height), but 3 ft
> generally works ok. Don't go too high--you'll
> generate pressure-induce artifacts. The only other
> comment I can make is that it takes practice (you
> know--how do you get to Carnegie Hall? practice,
> practice, practice). Oh, other details are coming up
> (it has been a while, so the files are dusty in my
> mental archives). It is best to prewarm your
> solutions--either by warming the bottles just prior to
> use or running coils of the tubing through a warm
> water bath (that may take a bit of experimenting to
> determine length of tubing, number of coils, water
> temp). The final temp should be close to 37C--this
> lessens vasoconstriction. An alternative is to use a
> vasodilator in the saline--something that most people
> prefer to avoid due to possible effects on vascular
> structures or antigens (I am an electron microscopist,
> primarily, so that is a very important consideration).
> The choice of fixative is wholly independent of the
> setup, and can be changed to fit the experiment (I
> have even used hydrogen sulfide laced fixative for
> metal localization--nasty, even in a hood!!!). Hope
> this helps.
>
> Roger Moretz, Ph.D.
> Dept of Toxicology
> Boehringer Ingelheim Pharmaceuticals, Inc.
>
> - --- "Rodriguez, Richard"
> <richard.rodriguez@spcorp.com> wrote:
> > Recently there were some interesting postings on rat
> > perfusions. We have
> > been trying mouse perfusions but have not been very
> > successful. Is there
> > anyone out there with experience with the mouse? We
> > would appreciate any
> > advice.
> >
> > Thanks
> >
> > Richard Rodriguez
> > Schering-Plough Research Institute
> > Lafayette, NJ
> > 973-940-4282
> >
> >
> ***************************************************************
> > This electronic message, including its attachments,
> > is confidential and
> > proprietary and is solely for the intended
> > recipient. If you are not the
> > intended recipient, this message was sent to you in
> > error and you are hereby
> > advised that any review, disclosure, copying,
> > distribution or use of this
> > message or any of the information included in this
> > message by you is
> > unauthorized and strictly prohibited. If you have
> > received this electronic
> > transmission in error, please immediately notify the
> > sender by reply to this
> > message and permanently delete all copies of this
> > message and its
> > attachments in your possession. Thank you.
> >
>
>
> __________________________________________________
> Do You Yahoo!?
> Yahoo! Auctions - buy the things you want at great prices
> http://auctions.yahoo.com/
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 09:07:07 -0500
> From: Hamish.Wilson@astrazeneca.com
> Subject: RE: Immunostainer
>
> Quite right, the Dako is made by LabVision - we have the LabVision version
> and are very happy with the machine and the service from LabVision UK.
>
>
>
> - -----Original Message-----
> From: Bonnie McMahill [mailto:bjmcm@mail.com]
> Sent: 01 May 2001 17:13
> To: 'Brennan, Liam'; HISTONET
> Subject: RE: Immunostainer
>
>
> We have two DAKO's at two facilities and love them (we previously had
> Ventana and BioTek). From what I understand though, the LabVision is a
> DAKO
> unit...(?)
>
> Good Luck,
>
> Bonnie McMahill
> Sacred Heart Medical Center
> Pathology Associates Inc, PS
> Spokane, WA
>
> - ------Original Message------
> From: "Horn, Hazel V" <HornHV@archildrens.org>
> To: "'Brennan, Liam'" <Liam.Brennan@bll.n-i.nhs.uk>, HISTONET
> <histonet@pathology.swmed.edu>
> Sent: April 30, 2001 2:42:44 PM GMT
> Subject: RE: Immunostainer
>
>
> I would go with DAKO...............I know it's not on your list....But,
> you
> should give it a try.
> Hazel
>
> > -----Original Message-----
> > From: Brennan, Liam [SMTP:Liam.Brennan@bll.n-i.nhs.uk]
> > Sent: Monday, April 30, 2001 6:02 AM
> > To: HISTONET
> > Subject: Immunostainer
> >
> > 30/04/01
> >
> > Anyone out there with an opinion/comment to make about either the
> Optimax
> > or
> > the Labvision immunostainer's as both companies have tendered to supply
> us
> > with an immunostainer as well Ventana.
> >
> > many thanks
> > Liam Brennan
> > Histopathology Dept.
> > Belfast City Hospital
> >
>
> ........................................................
> iWon.com http://www.iwon.com why wouldn't you?
> ........................................................
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 09:26:58 -0500
> From: "Dr. Allen A. Smith" <asmith@mail.barry.edu>
> Subject: Re: sirius red F3B
>
> Aldrich (1-800-558-9160) sells Sirius Red (C.I. 35780) under the name of
> "Direct Red 80" (catalog # 36-554-8,
> $13.60 for 5 grams).
>
> Allen A. Smith
> Barry University School of Graduate Medical Sciences
> Miami Shores, Florida
>
> - ----- Original Message -----
> From: Rebecca S Smith <bssvpisu@iastate.edu>
> To: HistoNet Server <histonet@pathology.swmed.edu>
> Sent: Monday, April 30, 2001 9:46 AM
> Subject: sirius red F3B
>
>
> > Help histonetters,
> > I have a scientist wanting me to do a stain called Sirius Red
> (Llewellyn,
> > 1970) I have found a procedure in Bancroft's book. Whew! Then I look
> in
> > I don't know how many catalogs and find Sirius Red F3B NOWHERE. I
> looked
> > in my old book of biological dyes for a possible alternate name. No
> > Luck! Does anyone know where I can get this stuff. AND once I find it
> any
> > clues. I have been warned that the preparation of this dye is tricky
> and
> > it can precipitate out on you quite easily. Thanks in advance!
> >
> >
> >
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 09:27:17 -0500
> From: Deb Boswell Lane <dbl@aretha.jax.org>
> Subject: BrdU in Unicryl??
>
> I'm relaying a question from a grad student:
> Limulus (horseshoe crab) was embedded in Unicryl and initial attempts at
> immunohistochemistry for BrdU haven't worked. Has anyone done BrdU immuno
> work for light microscopy in Unicryl or similar media? And does anyone
> have any suggestions for keeping 10 micron thick paraffin sections of
> Limulus (horseshoe crab) tissue on slides through IHC procedures? Please
> make suggestions! This if for a dissertation?
>
> Thanks in advance!
>
> Deb
> Deb Boswell Lane
> Professional Biomedical Technologist
> Biological Imaging
> The Jackson Laboratory
> 600 Main Street
> Bar Harbor, Maine 04609
> (207) 288-6193
> dbl@jax.org
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 09:27:39 -0500
> From: Gayle Callis <uvsgc@montana.edu>
> Subject: Rat CD3
>
> Serotec 1F4 clone, IgM istotype, and BD Pharmingen clone G4-18 (not
> recommended for paraffin)
>
> We grew up our own antibody from ATCC cell culture
> Gayle Callis
> Histopathology Supervisor
> Veterinary Molecular Biology
> Montana State University - Bozeman
> Bozeman MT 59717-3610
>
> 406 994-6367
> 404 994-4303 (FAX)
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 10:21:47 -0500
> From: "Alex Bolivar" <alexbolivar@hotmail.com>
> Subject: silver nitrate for marking margins
>
> <html><DIV>Hello all!</DIV>
> <DIV> </DIV>
> <DIV>I was wondering if anyone out there was using silver nitrate to mark
> the
> margins of gross specimens. We are currently using india ink
> and other
> tissue marking dyes, but would like to try silver nitrate. If anyone is
> using
> it, what concentration do you use, and do you have any concerns
> related
> to toxicity. Also, does it make a mess on cutting boards,
> instruments and things? </DIV>
> <DIV> </DIV>
> <DIV>Thanx in advance!</DIV>
> <DIV> </DIV>
> <DIV>Alex Bolivar</DIV>
> <DIV>University of Alberta Hospital</DIV>
> <DIV>Edmonton, Alberta</DIV>
> <DIV>Canada </DIV><br clear=all><hr>Get Your Private, Free E-mail
> from
> MSN Hotmail at <a
> href="http://www.hotmail.com">http://www.hotmail.com</a>.<br></p></html>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 10:38:14 -0500
> From: Lynn Gardner <lynn-gardner@uiowa.edu>
> Subject: Re: GEWF colon CA lymph node fixative
>
> Hey Bob,
>
> Just some information I though you might be interested in! This GEWF has
> been around for a while (may not be exact but is real close) we buy this
> commercially which is something called Pen-Fix, which is basically a
> modified alcoholic formalin. Alcoholic formalin has been around since
> Dezna
> Sheehan's time, however, not many people used straight alcoholic formalin
> due to it's drying effect on tissues like spleen, liver and uterus. The
> addition of the acetic acid seemed to make the difference.
>
> I have been using Pen-Fix for close to 20 years and here is what we do.
> You
> receive the speicmen in 10% NBF then trasfer to Pen-Fix when received in
> the lab (with the exception of small biopsies). Allow to sit overnight and
> the nodes just pop out at you, no more hunting! It also helps to sort of
> fix fatty tissues ( it doesn't truely fix the fat but removed the water
> making it more firm and easier to cut). This product when used with less
> time in the alcohol and xylene on the processor gives you beautiful
> morphology, increases the intensity of routine and special staining and is
> great for immunohistochemistry and In Situ.
>
> No I am not a sales rep for this product but it does work. I use this a
> lot
> in my work and so far have been extremely happy with it!
>
> Sincerely,
> Lynn Gardner, HT(ASCP)
> Senior Histological Technician
> The University of Iowa
> Anatomy and Cell Biology
> 51 Newton Road
> 1-100 BSB
> Iowa City, IA 52242
> Phone: 319-353-5490
> Fax: 319-335-7770
>
> At 06:48 PM 5/1/01 -0400, you wrote:
> >A lot of us on the Pathology (PATHO-L) and Histology (HistoNet)
> listservers
> >use special fixatives to facilitate retrieval of lymph nodes from
> colorectal
> >cancer specimens. A group from the University of Western Ontario (at
> London)
> >has an interesting method I'd like to try out. What's different about
> this
> >technique from others I've seen is the use of the special fixative as a
> >postfixative after overnight formalin fixation, a real convenience if you
>
> >receive a lot of half-fixed specimens such as most of us do.
> >
> >"GEWF solution: an inexpensive, simple, and effective aid for the
> retrieval
> >of lymph nodes from colorectal cancer specimens."
> >Ken J. Newell, Barry W. Sawka, Brian F. Rudrick, David K. Driman.
> >Arch Pathol Lab Med, May 2001;125:642-645.
> >
> >GEWF solution consists of:
> > 100 parts absolute ethanol [or reagent alcohol, I'm sure]
> > 34 parts water
> > 16 parts strong (37%) formalin
> > 10 parts glacial acetic acid
> >mixed in the order listed [why does that matter?]
> >
> >Open, pin out, and fix the specimen overnight in neutral buffered
> formalin
> >detach the pericolic fat
> >postfix it overnight in GEWF solution
> >dissect out lymph nodes
> >
> >The technique increased the average number of nodes recovered from 7 to
> 10,
> >doubled the number of positive nodes found, and decreased the average
> size of
>
> >positive nodes from 7 m to 5 mm. The authors found no interference with
> >immunohistochemical stains.
> >
> >Bob Richmond
> >Samurai Pathologist
> >Knoxville TN
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 11:14:03 -0500
> From: Bruce Gapinski <BGapinski@pathgroup.com>
> Subject: silver nitrate for marking margins
>
> Why silver? It's unnecessarily expensive! We use India ink, and Mrs.
> Stewarts Bluing.
> Bruce Gapinski HT(ASCP)
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 11:40:08 -0500
> From: Patsy.Ruegg@UCHSC.edu
> Subject: RE: BrdU in Unicryl??
>
> Katie Bennett of Lovelace Resp. res. Inst. Faxed me her procedure for
> brduin
> GMA (Immunobed). Don't have her email address handy but here is her phone
> number 505-845-1118 I think she is at Michigan State.
> Patsy Ruegg
>
> -----Original Message-----
> From: Deb Boswell Lane [mailto:dbl@aretha.jax.org]
> Sent: Wednesday, May 02, 2001 7:46 AM
> To: Histonet@pathology.swmed.edu
> Subject: BrdU in Unicryl??
>
> I'm relaying a question from a grad student:
> Limulus (horseshoe crab) was embedded in Unicryl and initial
> attempts at
> immunohistochemistry for BrdU haven't worked. Has anyone
> done BrdU immuno
> work for light microscopy in Unicryl or similar media? And
> does anyone
> have any suggestions for keeping 10 micron thick paraffin
> sections of
> Limulus (horseshoe crab) tissue on slides through IHC
> procedures? Please
> make suggestions! This if for a dissertation?
>
> Thanks in advance!
>
> Deb
> Deb Boswell Lane
> Professional Biomedical Technologist
> Biological Imaging
> The Jackson Laboratory
> 600 Main Street
> Bar Harbor, Maine 04609
> (207) 288-6193
> dbl@jax.org
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 11:40:24 -0500
> From: Patsy.Ruegg@UCHSC.edu
> Subject: RE: BrdU in Unicryl??
>
> A good way to keep sections on the slides if you are not doing collagen
> markers (the glue interfers with staining) is to precoat the slides (reg.
> Slides) with a solution of 5% elmers type glue in water, dip the slides in
> this then let them air dry. Pick up your sections onto the glue coated
> slides and dry them as usual, they stick really well, I use this for
> rabbit
> bones, but beware that glue is collagen based and will cause problems with
> some IHC markers.
> Patsy Ruegg
>
> -----Original Message-----
> From: Deb Boswell Lane [mailto:dbl@aretha.jax.org]
> Sent: Wednesday, May 02, 2001 7:46 AM
> To: Histonet@pathology.swmed.edu
> Subject: BrdU in Unicryl??
>
> I'm relaying a question from a grad student:
> Limulus (horseshoe crab) was embedded in Unicryl and initial
> attempts at
> immunohistochemistry for BrdU haven't worked. Has anyone
> done BrdU immuno
> work for light microscopy in Unicryl or similar media? And
> does anyone
> have any suggestions for keeping 10 micron thick paraffin
> sections of
> Limulus (horseshoe crab) tissue on slides through IHC
> procedures? Please
> make suggestions! This if for a dissertation?
>
> Thanks in advance!
>
> Deb
> Deb Boswell Lane
> Professional Biomedical Technologist
> Biological Imaging
> The Jackson Laboratory
> 600 Main Street
> Bar Harbor, Maine 04609
> (207) 288-6193
> dbl@jax.org
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 12:08:23 -0500
> From: "Aneeta Patel" <apatel@usuhs.mil>
> Subject: factor 8
>
> hi all
> I am trying to find out if anyone has done factor 8 immunostaining on
> parafiin
> embedded tissues and if so can you kindly share the details of method
> including ag retrival,thanks
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 12:08:40 -0500
> From: Shawn Griffin <SGriffin@ameripath.com>
> Subject: FW: FW: RCC
>
>
> Hey Richard
>
> One of our doctors went to a meeting and they talked about RCC. When she
> came back she had us order it. We have only used it a few times and she
> seems to be pleased with the results. Maybe you have found something out.
>
> Shawn
>
> - -----Original Message-----
> From: Richard Cartun [mailto:Rcartun@harthosp.org]
> Sent: Tuesday, May 01, 2001 10:40 AM
> To: SGriffin@ameripath.com
> Subject: Re: FW: RCC
>
>
> Hi Shawn:
>
> Do you find this antibody useful? It appears to lack sensitivity
> (identifying metastatic renal cell CA) from the work that I have done.
>
> R. Cartun
>
> >>> Shawn Griffin <SGriffin@ameripath.com> 04/27/01 11:49AM >>>
>
>
> - -----Original Message-----
> From: Shawn Griffin
> Sent: Friday, April 27, 2001 4:06 AM
> To: 'Patti Loykasek'
> Subject: RE: RCC
>
>
> Patti,
>
> We use RCC, clone 66.4c2 on paraffin tissue from NovoCastra. We pretreat
> the slides for 20 min using the steamer method. We use everything from
> Dako. The kit we are using is LSAB2HRP. The dilution is 1:100 for 30
> minutes. I hope this will help you some.
>
> Shawn Griffin
> sgriffin@ameripath.com
> Ameripath, Dallas
>
> - -----Original Message-----
> From: Patti Loykasek [mailto:ploykasek@phenopath.com]
> Sent: Thursday, April 26, 2001 5:11 PM
> To: histonet
> Subject: RCC
>
>
> Can anyone recommend a RCC clone that works on FFPE tissue? Pretreatments?
> We have tried several clones and pretreatments with inconsistent results.
> Thanks for the help.
>
> Patti Loykasek
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 12:11:40 -0500
> From: Nperson211@aol.com
> Subject: Osteonectin (SPARC) antibody..help!
>
> Histonetters,
> Does anyone use an osteonectin antibody reliably in formalin fixed,
> paraffin
> embedded rat tissues (brain)?
> If so, please share info and you will have my undying gratitude.
> Nancy Lemke
> Henry Ford Hospital
> Detroit, MI
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 12:15:11 -0500
> From: Patsy.Ruegg@UCHSC.edu
> Subject: RE: histology services
>
> Janet Maass sold her business to someone else and they sent me an email
> but
> I lost track of it, keep bugging me and I will see if I can get the new
> number for you.
> Patsy Ruegg
>
> -----Original Message-----
> From: Robert Geske [mailto:RGeske@lexgen.com]
> Sent: Wednesday, May 02, 2001 6:39 AM
> To: 'histonet@pathology.swmed.edu'
> Subject: histology services
>
> Does anyone know how to contact the histology service in
> Colorado that I
> believe is named Maas Histology. I would appreciate the
> phone number.
>
>
> thanks
>
> rob
>
> ****** The contents of this communication are intended only
> for the
> addressee and may contain confidential and/or privileged
> material. If you
> are not the intended recipient, please do not read, copy,
> use or disclose
> this communication and notify the sender. Opinions,
> conclusions and other
> information in this communication that do not relate to the
> official
> business of my company shall be understood as neither given
> nor endorsed by
> it. *****
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 12:15:34 -0500
> From: Abizar Lakdawalla <abizarl@innogenex.com>
> Subject: Elmer's glue
>
> I was under the impression that Elmer's white glue was collagen based!
> (embarassed) Abizar
> www.innogenex.com
>
> Patsy.Ruegg@UCHSC.edu wrote:
>
> > A good way to keep sections on the slides if you are not doing collagen
> > markers (the glue interfers with staining) is to precoat the slides
> (reg.
> > Slides) with a solution of 5% elmers type glue in water, dip the slides
> in
> > this then let them air dry. Pick up your sections onto the glue coated
> > slides and dry them as usual, they stick really well, I use this for
> rabbit
> > bones, but beware that glue is collagen based and will cause problems
> with
> > some IHC markers.
> > Patsy Ruegg
> >
> > -----Original Message-----
> > From: Deb Boswell Lane [mailto:dbl@aretha.jax.org]
> > Sent: Wednesday, May 02, 2001 7:46 AM
> > To: Histonet@pathology.swmed.edu
> > Subject: BrdU in Unicryl??
> >
> > I'm relaying a question from a grad student:
> > Limulus (horseshoe crab) was embedded in Unicryl and
> initial
> > attempts at
> > immunohistochemistry for BrdU haven't worked. Has
> anyone
> > done BrdU immuno
> > work for light microscopy in Unicryl or similar media?
> And
> > does anyone
> > have any suggestions for keeping 10 micron thick
> paraffin
> > sections of
> > Limulus (horseshoe crab) tissue on slides through IHC
> > procedures? Please
> > make suggestions! This if for a dissertation?
> >
> > Thanks in advance!
> >
> > Deb
> > Deb Boswell Lane
> > Professional Biomedical Technologist
> > Biological Imaging
> > The Jackson Laboratory
> > 600 Main Street
> > Bar Harbor, Maine 04609
> > (207) 288-6193
> > dbl@jax.org
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 12:15:49 -0500
> From: Eileen_Dusek@cdh.org
> Subject: Age related Joint Commession question
>
> Recently someone wrote about an age related question for the Joint
> Commission. I was just asked to see if this would be pertinent to
> Histology. Does anyone have a clue
>
> Thanks
>
>
> Eileen
> Central Dupage Hospital.
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 13:47:02 -0500
> From: Gayle Callis <uvsgc@montana.edu>
> Subject: Attn: Patsy Ruegg
>
> Patsy,
>
> For some reason there is a circular reference with an alias in your
> address, and I cannot reply to you directly, no one elses email addresses
> do this to me, so this is coming through HIstonet, sorry folks! gremlins
> in the computer dancing on the keys, messing up address files or some
> little thingee ma bob!
>
>
> Sigma Dulbeccos PBS, D 5652 cat #, makes up 50 liters. Make sure you get
> the one without magnesium or calcium.
>
> I make up my own TRIS buffered saline (TBS), you can find a recipe for 10X
> TBS on website, eBioscience.com, just dilute and run with it. I use it at
> ph 7.6, and for AP protocols.
> Gayle Callis
> Histopathology Supervisor
> Veterinary Molecular Biology
> Montana State University - Bozeman
> Bozeman MT 59717-3610
>
> 406 994-6367
> 404 994-4303 (FAX)
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 14:03:01 -0500
> From: brian.chelack@usask.ca
> Subject: Ficin comments
>
> Hello Shannon;
>
> I have used ficin in the distant past and found it to be a
> very weak enzyme, compared to pronase or trypsin. It was if
> I remember correctly it was used mainly by the blood banking
> industry. I have an older reference which may of use to you
> in the comparison of various enzymatic treatments;
>
> Towle, A.C., Lauder, J.M., and Joh, T.H., Optimization of
> tyrosinase hydroxylase immunocytochemistry in poaraffin
> sections using pretreatment with proteolytic enzymes, J.
> Histochem. Cytochem., 32,766,1984.
>
> regards
>
> Brian Chelack
> Prairie Diagnostic Services
> Saskatoon
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 14:03:17 -0500
> From: Bruce Gapinski <BGapinski@pathgroup.com>
> Subject: RE: silver nitrate for marking margins
>
> Any grocery store here on the West Coast has it. Can't believe it but they
> actually have a web address http://www.mrsstewart.com . It's made byLuther
> Ford & Co. P.O.Box 201405 Bloomington, MN 55420.
> Bruce Gapinski HT(ASCP)
>
> -----Original Message-----
> From: Jonathan R. Oppenheimer
> [mailto:joppenheimer@ourlab.net]
> Sent: Wednesday, May 02, 2001 10:20 AM
> To: Bruce Gapinski
> Subject: RE: silver nitrate for marking margins
>
> Do you know where I can get Mrs. Stewarts Bluing ?
>
> -----Original Message-----
> From: Bruce Gapinski [mailto:BGapinski@pathgroup.com]
> Sent: Wednesday, May 02, 2001 10:49 AM
> To: 'HistoNet@Pathology.swmed.edu'
> Subject: silver nitrate for marking margins
>
>
> Why silver? It's unnecessarily expensive! We use India ink,
> and Mrs.
> Stewarts Bluing.
> Bruce Gapinski HT(ASCP)
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 14:03:35 -0500
> From: "Kim Kusser" <kkusser@trudeauinstitute.org>
> Subject: Re:PNA staining
>
> Hi,
>
> I stain all the time with PNA on mouse spleen. I suppose I can just tell
> you
> what I do:
>
>
> 1. cut frozen sections 5-7um-----> air dry (1hr to O/N)
> 2. Fix in 4c acetone for 10 minutes -----> air dry ---> PBS
> 3. Block endogenous peroxidase with 0.1M NaN3 for 10 minutes ----> PBS 3X
> (3x
> 5 minutes)
> 4. Serum block for 30 minutes (I use 5%BSA in dPBS)
> 5. Avidin block 15 minutes ----> pBS1X ----> Biotin block 15 minutes ---->
> PBS
> 2X
> 6. PNA-b 1:800 for 30 minutes ------> PBS 3X
> 7. ABC (from vector) for 30 minutes ----> PBS 3X
> 8. Develope with DAB
>
> PNA is very sticky, so you'll want to make sure you titrate it as high as
> you
> can. But I routinely get very nice germinal center staining.
>
> Hope this helps.
>
> Kim Kusser
>
>
>
>
>
> Date: 1 May 2001 16:49:33 -0500
> From: Jianyun.Yin@aventis.com
> Subject: PNA staining
>
> Dear all,
>
> We are having difficulty get nice PNA (for Germinal center B cells)
> staining on mouse spleen and LN. Can any one share some experience about
> this staining with us? Thanks!
>
> Jian Yin
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 14:03:52 -0500
> From: Patsy.Ruegg@UCHSC.edu
> Subject: RE: Osteonectin (SPARC) antibody..help!
>
> I use osteonectin from U of Iowa Developmental Studies Hybridoma Bank on
> rat
> tissues, mainly bone and cartilage ffpe, formic acid decaled,
> pre-treatment
> is pepsin digestion for 10 min. at 37 dC.
> Patsy Ruegg
>
> -----Original Message-----
> From: Nperson211@aol.com [mailto:Nperson211@aol.com]
> Sent: Wednesday, May 02, 2001 10:35 AM
> To: histonet@pathology.swmed.edu
> Subject: Osteonectin (SPARC) antibody..help!
>
> Histonetters,
> Does anyone use an osteonectin antibody reliably in formalin
> fixed, paraffin embedded rat tissues (brain)?
> If so, please share info and you will have my undying
> gratitude.
> Nancy Lemke
> Henry Ford Hospital
> Detroit, MI
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 14:04:43 -0500
> From: "Kopczynski, Charlotte" <Charlotte.Kopczynski@baycare.org>
> Subject: RE: Age related Joint Commession question
>
> It is my understanding that the JCAHO standard relates to direct patient
> contact. Histology would not be involved unless histotechs are answering
> the phone or coming into contact with patients. There is age specific
> training whcih must be documented. In my laboratory, only the clerks in
> the
> Pathology office are required to take this training. If this is not
> correct, please let me know...
> Charlotte Kopczynski
> Technical Histology Manager
> Baycare Laboratory Services
> Phone: 727-461-8246
> Fax: 727-462-7617
>
>
>
> > -----Original Message-----
> > From: Eileen_Dusek@cdh.org [SMTP:Eileen_Dusek@cdh.org]
> > Sent: Tuesday, May 01, 2001 4:11 PM
> > To: HistoNet@pathology.swmed.edu
> > Subject: Age related Joint Commession question
> >
> > Recently someone wrote about an age related question for the Joint
> > Commission. I was just asked to see if this would be pertinent to
> > Histology. Does anyone have a clue
> >
> > Thanks
> >
> >
> > Eileen
> > Central Dupage Hospital.
> >
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 14:05:11 -0500
> From: "Jamie Erickson" <JErickson@genetics.com>
> Subject: Re: factor 8
>
> Hi All,
> I work with a rabbit anti- human VWF antibody (factor VIII)
> (DAKO)
> at 3.8ug/mL in 10% NBF fixed(24 hours) paraffin embedded mouse tissues
> Prior
> to staining, the paraffin-embedded slides for vWF staining were enzyme
> digested with 0.5% Pronase respectively for 15 minutes at 37 #161# C.
> This is
> done using a peroxidase based IHC system.
> That being said it would be Very Very Very helpful if people would but
> specify species they are interested in so that this information is not a
> was
> of time....Mouse, Rat, Human, Monkey we are not all in a Clinical
> Research....
> Thanks
>
>
>
> Jamie Erickson
> Scientist II
> Genetics Institute
> 1 Burtt Rd.
> Andover, MA 01810
> work : (978) 247-1348
> FAX : (978) 247-1389
>
> >>> Aneeta Patel <apatel@usuhs.mil> 05/02 12:21 PM >>>
> hi all
> I am trying to find out if anyone has done factor 8 immunostaining on
> parafiin
> embedded tissues and if so can you kindly share the details of method
> including ag retrival,thanks
>
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 14:05:44 -0500
> From: nlgervais@mmm.com
> Subject: Feedback Stat
>
> Hello,
> I am looking for the login name and password for the Feedback Stat
> NSH
> survey. Thanks in advance.
>
>
> Nicole Gervais, B.S.
> 3M Pharmaceuticals
> St. Paul, MN
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 14:05:58 -0500
> From: "Histopathology Lab" <histopathology@cvm.msstate.edu>
> Subject: CAP Workload Values
>
> Can anyone tell me the workload value assigned to IHC Procedures? I
> am at
> a Vet School Lab and of we are not under CAP regs. However, our Laboratory
> Manager does loosely use the CAP System to figure some charges and
> staffing . Thanks
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 16:18:51 -0500
> From: "amdj@duke.edu" <amdj@duke.edu>
> Subject: RE: Age related Joint Commission question
>
> If you have patient contact with patients of varying ages (particularly
> but
> not necessarily limited to pediatric and geriatric patients) OR if your
> work (e.g. procedures, laboratory reference ranges, alert values) varies
> according to patient age the Joint Commission quite reasonably requires
> than your competency testing specifically address differences in the
> procedures you may have to follow for patients of different ages. Judging
> from at least one posting I saw earlier there are some "managers"
> requiring
> people to invent stuff so that everyone in the hospital will have age
> specific competencies. If you have no procedures that vary according to
> the
> age of the patient and no patient contact, you probably can check this
> "does not apply". This lets the vast majority of histotechs off the
> hook--but not all.
>
> Bert
>
> - -----Original Message-----
> From: Eileen_Dusek@cdh.org [SMTP:Eileen_Dusek@cdh.org]
> Sent: Tuesday, May 01, 2001 4:11 PM
> To: HistoNet@pathology.swmed.edu
> Subject: Age related Joint Commession question
>
> Recently someone wrote about an age related question for the Joint
> Commission. I was just asked to see if this would be pertinent to
> Histology. Does anyone have a clue
>
> Thanks
>
>
> Eileen
> Central Dupage Hospital.
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 16:19:42 -0500
> From: "Vinnie Della Speranza" <dellav@musc.edu>
> Subject: Re: CAP Workload Values
>
> If you are referring to the CAP Workload Recording System, this program
> was
> abandoned in 1992.
> However, at that time, they alloted 13 minutes per slide for a two layer
> (indirect) ihc stain, 8.0 minutes per slide for a one layer (or direct)
> method
> performed manually (non-automated). There was no allocation for a three
> layer
> method ( i.e, primary ab plus secondary ab conjugated with biotin and then
> avidin-HRP, an one example). I don't know how, if at all, this compares to
> what you may be doingn in your lab in 2001.
>
> More importantly, you might encourage your manager to consider
> participating
> in a benchmarking program, the results of which will allow you compare
> your
> operation to others of comparable complexity and size. This data is apt to
> be
> viewed as more credible to higher administration than the use of data from
> a
> system that was abandoned nine years ago.
>
>
>
> 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
>
>
> >>> Histopathology Lab <histopathology@cvm.msstate.edu> 05/02/01 02:54PM
> >>>
> Can anyone tell me the workload value assigned to IHC Procedures? I
> am at
> a Vet School Lab and of we are not under CAP regs. However, our Laboratory
> Manager does loosely use the CAP System to figure some charges and
> staffing . Thanks
>
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 16:19:59 -0500
> From: "John Ryan" <ZJPR01@sleh.com>
> Subject: Histology lab in Colorado
>
> This message is in response to a request by Robert Genske who was
> inquiring
> about a histology laboratory in Colorado by the name of Maass Histology.
> This
> lab was sold by Janet Maass over a year and a half ago and she is no
> longer
> affiliated with this lab in any capacity.
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 16:20:18 -0500
> From: Patricia Bourne <p_bourne_14526@yahoo.com>
> Subject: Re: Immunostainer
>
> What is the DAKO horizon...the Dako machines that we
> are using are the same as Lab Vision in fact they are
> made by lab vision...they do not require special slides.....
>
> __________________________________________________
> Do You Yahoo!?
> Yahoo! Auctions - buy the things you want at great prices
> http://auctions.yahoo.com/
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 16:28:20 -0500
> From: "In C. Kim" <inkim@insitus.com>
> Subject: Re: Elmer's glue
>
> If my memory serves correct, the main component of Elmer's glue is
> polyvinyl alcohol.
> Could someone correct me if I am wrong?
>
> In Kim
>
>
>
> - ----- Original Message -----
> From: "Abizar Lakdawalla" <abizarl@innogenex.com>
> To: <Patsy.Ruegg@UCHSC.edu>
> Cc: <dbl@aretha.jax.org>; <Histonet@pathology.swmed.edu>
> Sent: Wednesday, May 02, 2001 10:53 AM
> Subject: Elmer's glue
>
>
> > I was under the impression that Elmer's white glue was collagen based!
> > (embarassed) Abizar
> > www.innogenex.com
> >
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 16:28:52 -0500
> From: "Amos Brooks" <amosbrooks@home.com>
> Subject: Re: factor 8
>
> Hi,
> Our antibody (from DAKO) is kind of old so it wouldn't surprise me at
> all if you get a different dilution than we have now. After peroxidase
> blocking, we pretreat with tripsin for 15 min, then incubate for 30 min at
> room temp. at 1:10,000. (The dilution is why the antibody is old, it will
> last forever at this rate) Our detection kit is LSAB+ and DAB chromogen.
> hope this helps
> Amos Brooks
> - ----- Original Message -----
> From: "Aneeta Patel" <apatel@usuhs.mil>
> To: <Histonet@pathology.swmed.edu>
> Sent: Wednesday, May 02, 2001 12:21 PM
> Subject: factor 8
>
>
> hi all
> I am trying to find out if anyone has done factor 8 immunostaining on
> parafiin embedded tissues and if so can you kindly share the details of
> method including ag retrival,thanks
>
>
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 18:24:13 -0500
> From: "Morken, Tim" <tim9@cdc.gov>
> Subject: RE: CAP Workload Values
>
> The CAP assigned 13.0 units to each slide stained. This included the time
> spent manually handling the slide while doing prep work (AFTER sectioning)
> and staining. note that there was no AR in those days, so it didn't
> include
> that time. It made no allowance for automated immunos either.
>
> FYI, the CAP workoad method had a mechanism to determine your own units.
> It
> involved doing time studies. I don't have the books anymore, so I can't
> say
> more than that about it.
>
> I went to using raw numbers of hard items to count, rather than time
> units.
> It just became too problematic with all the variations on procedures. by
> hard items I mean actual numbers of cases, blocks, slides, stains, etc.
> Things that no one can argue about. I found that very effective in proving
> your workload.
>
> Tim Morken
> Atlanta
>
>
>
> histopathology@cvm.msstate.edu
> - -----Original Message-----
> From: Histopathology Lab [mailto:histopathology@cvm.msstate.edu]
> Sent: Wednesday, May 02, 2001 2:55 PM
> To: Histonet@pathology.swmed.edu
> Subject: CAP Workload Values
>
>
> Can anyone tell me the workload value assigned to IHC Procedures? I
> am at
> a Vet School Lab and of we are not under CAP regs. However, our Laboratory
> Manager does loosely use the CAP System to figure some charges and
> staffing . Thanks
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 18:24:30 -0500
> From: "amdj@duke.edu" <amdj@duke.edu>
> Subject: RE: CAP Workload Values
>
> This doesn't have a specific code, but rather a suffix .757 ( fell under
> the Immunology section and a different code was used for each antibody) at
>
> 13 minutes per slide not including sectioning which gets 88365 at 2
> minutes.
>
> Bert
>
> - -----Original Message-----
> From: Histopathology Lab [SMTP:histopathology@cvm.msstate.edu]
> Sent: Wednesday, May 02, 2001 2:55 PM
> To: Histonet@pathology.swmed.edu
> Subject: CAP Workload Values
>
> Can anyone tell me the workload value assigned to IHC Procedures? I
> am at
> a Vet School Lab and of we are not under CAP regs. However, our Laboratory
> Manager does loosely use the CAP System to figure some charges and
> staffing . Thanks
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 18:24:50 -0500
> From: Karen Larison <larisonk@uoneuro.uoregon.edu>
> Subject: Re: detergents in antibody diuents
>
> Ron,
>
> For detergents to be effective, they should be used at their critical
> micelle concentration. This is the concentration at which the
> detergent forms spherical aggregates with the fatty tails inside the
> sphere and the hydrophilic portion on the outside. The Sigma catalog
> provides a table of critical micelle concentrations in millimolar
> concentrations. You'll have to translate these into % concentrations.
>
> Karen in Oregon
>
>
>
>
>
> >I have seen various concentrations of detergents (Triton X, Tween 20
> >etc.) used in primary antibody diluents and wondered if there were any
> >guidelines for choosing a particular concentration. Does it depend on
> >whether paraffin or frozen sections are used or is it more dependent
> >upon the type and concentration of fixative? Also, is there an effect of
> >detergent concentration on the ability of sections to adhere to slides?
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 18:25:05 -0500
> From: "Wellik, Linda E." <Wellik.Linda@mayo.edu>
> Subject: RE: unsubscribe
>
>
>
>
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 18:26:14 -0500
> From: Denise Bland-Piontek <bland008@umn.edu>
> Subject: Chromogenic in situ staining with Y-probe
>
> Hello all:
> Is anyone out there doing any chromogenic in situ staining for the Y
> chromosome? I'm getting weak staining and would like any advice from
> others using this procedure. I'm using an m-RNA procedure from Biogenex
> and I've purchased a biotin labeled Y-probe from ID Labs. Currently I'm
> getting variable staining and would appreciate comparing the recipe I
> was given from the company with anyone out there who has been doing this
> on a regular basis. Thanks in advance,
> Denise Bland-Piontek, HTL (ASCP)
> University of Minnesota
> Anatomic Pathology Research Lab
>
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 18:26:37 -0500
> From: conmac@cc.usu.edu
> Subject: Re: detergents in antibody diuents
>
> I use tween 20 with TRIS buffer for all my rinses (IHC). I've never had
> a problem with sections coming off, however, I find that the surface
> tension on the slide is nill and unless I do a plain TRIS rinse, all the
> reagent runs off the slide.
>
> I use the tween 20 because i was told that it helps remove the excess
> reagent so you get cleaner rinsing between reagents. At one time, I
> used only plain TRIS buffer to rinse in, however, there was some
> nonspecific background staining that was hard to interpret. Doing the
> same number of rinses as before, the nonspecific background staining is
> gone since using the tween 20.
>
> Connie McManus
>
> Neuropathology wrote:
> >
> > We have used Tween 20 in our antibody diluent at a concentration of
> 0.05%
> > without any problems on both paraffin and frozen sections.
> >
> > Bob Quilty
> > Dept of Neuropathology
> > Frenchay Hospital
> > Bristol UK
> >
> > ----- Original Message -----
> > From: Ron Salisbury <ronald2@uakron.edu>
> > To: Histonet <histonet@pathology.swmed.edu>
> > Sent: Tuesday, May 01, 2001 7:06 PM
> > Subject: detergents in antibody diuents
> >
> > > I have seen various concentrations of detergents (Triton X, Tween 20
> > > etc.) used in primary antibody diluents and wondered if there were any
> > > guidelines for choosing a particular concentration. Does it depend on
> > > whether paraffin or frozen sections are used or is it more dependent
> > > upon the type and concentration of fixative? Also, is there an effect
> of
> > > detergent concentration on the ability of sections to adhere to
> slides?
> > >
> > >
>
> - --
> D#236##017#a#193##177##026#a
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 22:19:43 -0500
> From: Nperson211@aol.com
> Subject: anti human osteonectin
>
>
> I will try this again. I am staining human tumors implanted into rat
> brains,
> so I need info on an anti human osteonectin. I have been inhaling
> paraffin
> fumes for too many years and the miserable effects are starting to show.
> Again, my gratitude to all who reply, even to commiserate with my confused
>
> state.
> Thanks,
> Nancy Lemke
> Henry Ford Hospital
> Detroit, MI
>
>
>
> ******************* NOTE *******************
> There may be important message content
> contained in the following MIME Information.
> ********************************************
>
>
> - ------------------ MIME Information follows ------------------
>
>
> - --part1_b7.dc4cc44.282226a6_boundary
> Content-Type: text/plain; charset="US-ASCII"
> Content-Transfer-Encoding: 7bit
>
> <<<<<< See above "Message Body" >>>>>>
>
> - --part1_b7.dc4cc44.282226a6_boundary
> Content-Type: text/html; charset="US-ASCII"
> Content-Transfer-Encoding: 7bit
>
> <HTML><FONT FACE=arial,helvetica><FONT SIZE=2>I will try this again.
> I
> am staining human tumors implanted into rat brains,
> <BR>so I need info on an anti human osteonectin. I have been
> inhaling
> paraffin
> <BR>fumes for too many years and the miserable effects are starting to
> show.
> <BR>Again, my gratitude to all who reply, even to commiserate with my
> confused
>
> <BR>state.
> <BR>Thanks,
> <BR>Nancy Lemke
> <BR>Henry Ford Hospital
> <BR>Detroit, MI</FONT></HTML>
>
> - --part1_b7.dc4cc44.282226a6_boundary--
>
>
> ----------------------------------------------------------------------
>
> Date: 2 May 2001 23:45:27 -0500
> From: "J. A. Kiernan" <jkiernan@uwo.ca>
> Subject: Re: Elmer's glue: Reflexions and Quexions.
>
> (Pertinent bits of the emails that prompted these comments
> are cited with > >> etc at the end of my two screenfuls.)
>
> Traditional glue is certainly collagen based. It's very concentrated
> gelatin, made by prolonged boiling of bones, and has to be heated
> to reduce its viscosity to a workable level. I think it's still used
> industrially, for jobs like sticking the layers together in plywood.
> The dried and hardened glue can be permanently cross-linked by
> exposure to formaldehyde gas.
>
> In modern times the word glue is applied to all sorts of adhesives.
> The impression I've been under for many years is that the white
> ones like Elmer's have polyvinyl acetate as the main ingredient.
> In 1983 two Finns (Jarvinen and Rinne, Acta Histochem. 72:751-752)
> wrote a short paper describing the use of a polyvinyl acetate
> adhesive for sections. To those of us who requested reprints
> they enclosed a small green squeezy-tube with a trade name on
> it, and also some other printing which, when examined with a
> microscope, turned out to be entirely in Finnish.
>
> (The Finns developed the micro-printing technology to accommodate
> statements in a language that has very few short words. In earlier
> times they could produce only large tubes of glue, toothpaste etc.
> For 2-gram tubes of Finnish eye ointment you now need an oil immersion
> objective to read the instructions, but it works so well that after
> finishing the treatment a X40 high-dry is sufficient.)
>
> The stuff that came out of the green tube looked, smelt and felt
> exactly like any other general-purpose white adhesive.
> I tried it a few times (the published paper was in English) but
> it was a messy technique and didn't seem to me at the time to be
> as good as chrome-gelatin.
>
> Patsy's Ruegg's procedure (quoted below) seems to be nothing like as
> messy as the one in the 1983 Acta Histochem paper. It is important,
> however, to know what's in Elmer's "glue" before using it as a
> section adhesive in research or diagnostic applications. Has anyone
> asked Elmer? Is there a publication that documents its introduction
> into microtechnique? Are the products of other major manufacturers
> similarly used? Wotta lotta questions.
> - ----------------------------------------
> John A. Kiernan
> Department of Anatomy & Cell Biology
> The University of Western Ontario
> London, Canada N6A 5C1
> kiernan@uwo.ca
> http://publish.uwo.ca/~jkiernan
> - --------------------------------------
> On Wed, 2 May 2001, Abizar Lakdawalla wrote:
> > I was under the impression that Elmer's white glue was collagen based!
> >
> > Patsy.Ruegg@UCHSC.edu had written:
> >
> > > A good way to keep sections on the slides if you are not doing
> collagen
> > > markers (the glue interfers with staining) is to precoat the slides
> (reg.
> > > Slides) with a solution of 5% elmers type glue in water, dip the
> slides in
> > > this then let them air dry. ...
> > > ... but beware that glue is collagen based and will cause problems
> > > with some IHC markers.
> - -------------------------------------------------
>
>
>
> Here are the messages received yesterday!
>
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