Help!  Does anyone out there in Histo Land know of a written guide for
first time presentors?  I am contemplating giving a short lecture in Nov
2000 at the annual Mohs conference in Denver.  Save for extreme stage
fright and no experience speaking to large groups of people I would just
plough through this challenge except I am one of those types who is never
too prepared for anything.  Any suggestions, ideas, etc would be
appreciated. Also how does one go about making overheads from notes?
Thanks in advance.

Sue Becker, HTL
Albany, NY

----------
> From: HistoNet Server <histonet@pathology.swmed.edu>
> To: HistoNet Server <histonet@pathology.swmed.edu>
> Subject: Daily Digest
> Date: Saturday, November 13, 1999 0:05 AM
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 03:15:29 -0600
> From: Mary Latimer <ml4@st-andrews.ac.uk>
> Subject: Re: Need Antigen Retrieval Help
>
> Dear Katie  I have no personel experience of this technique but on an
> immuno course That I attended in London  at the Hammersmith Hospital
when
> these techniques were being discussed one of the techs there said that
she
> did these in an autoclave rather than microwave her reasons were that the
> microwave needed constant watching and the addition of near boiling
> solution to avoid drying out this apparently is not the case if you use
an
> autoclave or pressure cooker there is a written method in a book called
> Introduction to Immunocytochemistry by Polak and Van Noorden I can fax it
> if you would like to see it please email ....Hope this Helps Mary
>
> On Thu, 11 Nov 1999, Bennett, Catherine (Katie) wrote:
>
> > Hi all!
> >
> > I'm starting a double staining IHC run this morning that is intending
to
> > take two days, and I may have already committed a fatal error on step
one,
> > so I'm wondering if I should continue or start over.
> >
> > I am doing microwave antigen retrieval and had a problem with the
solution
> > boiling over and completely out of the coplan jar.  My protocol calls
for
> > microvaving slides in a coplan jar of antigen retrieval for 2x 5 min,
but
> > after just 1.5 min all the solution is boiled over and gone.  Needless
to
> > say, I didn't catch it in time and the slides may have gone through
some
> > microwaving without any solution in the jar.  I had placed the coplan
jar in
> > a small dish of shallow water, so the microwave was very humid at
least.
> >
> > I'm guessing the microwave is much more powerful than the one used in
the
> > protocol I am using and I should experiment with which power setting I
> > should use.  However, in the meantime, are the slides from today toast
> > because they have been "cooked" without being in the antigen retrieval
> > solution, or can I continue with them?
> >
> > Any suggestions from others on preventing boil-over would also be much
> > appreciated.
> >
> > *********************************
> > Catherine "Katie" Bresee Bennett
> > Sr. Technical Associate
> > Lovelace Respiratory Research Institute
> > Albuquerque, New Mexico
> >
> >
> >
>
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 06:45:49 -0600
> From: Barbara Wright <bwright@gene.com>
> Subject: Re: Rat B lymphocyte marker
>
> Cindy;
>
> Pharmingen has a mouse anti-rat CD45R  (Clone HIS24) that they state
works in
> paraffin embedded
> tissues.
>
> Thanks
> Barb:)
>
> Cindy Farman wrote:
>
> > Hi All,
> >
> > Can anyone recommend an antibody that will mark B lymphocytes in
> paraffin-embedded rat tissue?
> >
> > Thank you,
> >
> > Cindy Farman
> > Sierra Biomedical, Inc.
>
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 06:46:16 -0600
> From: Barbara Wright <bwright@gene.com>
> Subject: Re: CD31,CD34
>
> Lee;
>
> huCD34 does not cross react with mouse.  Pharmingen has a rat anti-mouse
CD34
> (Clone RAM34) that works on frozen sections and a rat anti-mouse CD31
(Clone
> MEC13.3) that works on both frozen and paraffin mouse tissues. muCD31
works on
> 10%NBF, PLP and 4% Paraformaldehyde fixed paraffin embedded mouse kidney
with
> high ph citra (AR10 from Biogenex) antigen retrieval with tyramide
> amplification
> (TSA from NEN).  For most CD markers they are
> specific for the species intended especially if they are monoclonal
> antibodies.
> You may have some luck with with cross reactivity with polyclonals (e.g..
> rabbit
> anti-human CD3 does work on rodents).
> Thanks
> Barb:)
>
> Lee Ann Baldridge wrote:
>
> > Hi All,
> > I'm having big time trouble getting CD34 to stain in mouse kidney.  I
used
> Dako's primary and Vector's MOM. No staining. Also no staining with the
> Pharmingen's CD31(rat anti-mouse) in tumor cells implanted in a mouse
brain.
> I tried Dako's LSAB2 kit and Vector's MOM just for kicks.  Any help or
> suggestions would be greatly appriecated. I'm new at this animal tissue
stuff
> so be patient with my frequent questions. Thanks for your help.
> > Lee Ann Baldridge
> > I.U. Hosp.
> > Indpls., IN.
> > 317-274-5790
> > 317-278-4820 fax
>
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 08:00:44 -0600
> From: "Berger, Jennifer" <jberger@lrri.org>
> Subject: RE: Need Antigen Retrieval Help
>
> The way that we do it here with great results is to fill the coplin jar
with
> the antigen retrieval solution, place saran wrap over the jar secured
with a
> rubberband and place the whole thing into another container such as a
large
> beaker filled with water about 2/3 the way to the top of the coplin jar
and
> microwave.  I have gone for 10 min straight without any problems and with
> beautiful results.
> Jennifer
>
> Jennifer Berger
> Senior Technical Associate
> Lovelace Respiratory Research Institute
> Albuquerque, NM
> (505)845-1122; (505)845-1225
> jberger@lrri.org
>
> > -----Original Message-----
> > From:	ThaxtonPM@aol.com [SMTP:ThaxtonPM@aol.com]
> > Sent:	Thursday, November 11, 1999 8:00 PM
> > To:	cbennett@lrri.org; histonet@pathology.swmed.edu
> > Subject:	Re: Need Antigen Retrieval Help
> >
> > Hi Catherine,
> >   The easiest way to microwave AR is to watch the coplin jar while
> > microwaving on high heat. When you see the solution bubble, stop the
> > microwave and check the temperature (also record the time that it took
to
> > get
> > to temp). Microwave again on medium heat, and check the temperature at
> > each
> > interval to make sure the temperature is holding. It is best to do this
> > using
> > water to begin with, and always make sure you have the same number of
> > slides
> > in the coplin jar each time.
> > Once you get the procedure standardized with the microwave you are
using,
> > you
> > should be ok.
> >
> >
> > okay, back in my corner now,
> >         Phyllis
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 08:30:10 -0600
> From: Cynthia Favara <cfavara@niaid.nih.gov>
> Subject: RE: DAB: impermanent?
>
> Have also had DAB disappear glad to know it happens elsewhere and i ma
not
> totally crazy - just mostly
> Cynthia Favara
> NIAID/RML/LPVD
> 903 South 4th Street
> Hamilton, MT 59840
> PH: 406-363-9317
> FAX: 406-363-9286
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 09:15:47 -0600
> From: "Hugo M. Fernandez Bellon" <ivha23@blues.uab.es>
> Subject: Re: Nucleated red blood cells
>
> I have no idea on their lifespan. I always viewed them as a step previous
to
> non-nucleated RBC's (which only mammals have). As far as I can figure,
they
> are less effective in carrying oxygen (part of their volume is occupied
by
> the nucleus) and are less flexible (thus more fragile) and offer more
> resistance to flow than mammal RBC's.
>
>
> Siyami Karahan ha escrito:
>
> >  Dear histonetters:
> >
> >  Does anyone know what are the life spans of red blood cells in birds
and
> > fish? what kind of advantages does a nucleadted blood cell have?
> >
> >  Thank you
> >
> >  Siyami
>
>
>
>
> - --
> Hugo Fernandez < ivha23@blues.uab.es >
>
> U.D. Histologia i Anatomia Patologica
> Facultat de Veterinaria
> Universitat Autonoma de Barcelona
> 08193 Cerdanyola del Vallis
> ESPAQA
>
> Tel. 93 581 31 45
> Fax. 93 581 20 06
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 09:16:16 -0600
> From: emaher@zeiss.com
> Subject: Re: Need Antigen Retrieval Help
>
>      There is a good article by M.A. Hayat in the September issue of
>      Microscopy Today which explains all the aspects of antigen retrival
in
>      a microwave.
>      Evanne Maher
>      Microm Instruments/RAS
>
>
> ______________________________ Reply Separator
> _________________________________
> Subject: Need Antigen Retrieval Help
> Author:  "Bennett; Catherine (Katie)" <cbennett@lrri.org> at Internet
> Date:    11/11/99 9:20 AM
>
>
> Hi all!
>
> I'm starting a double staining IHC run this morning that is intending to
> take two days, and I may have already committed a fatal error on step
one,
> so I'm wondering if I should continue or start over.
>
> I am doing microwave antigen retrieval and had a problem with the
solution
> boiling over and completely out of the coplan jar.  My protocol calls for
> microvaving slides in a coplan jar of antigen retrieval for 2x 5 min, but
> after just 1.5 min all the solution is boiled over and gone.  Needless to
> say, I didn't catch it in time and the slides may have gone through some
> microwaving without any solution in the jar.  I had placed the coplan jar
in
> a small dish of shallow water, so the microwave was very humid at least.
>
> I'm guessing the microwave is much more powerful than the one used in the
> protocol I am using and I should experiment with which power setting I
> should use.  However, in the meantime, are the slides from today toast
> because they have been "cooked" without being in the antigen retrieval
> solution, or can I continue with them?
>
> Any suggestions from others on preventing boil-over would also be much
> appreciated.
>
> *********************************
> Catherine "Katie" Bresee Bennett
> Sr. Technical Associate
> Lovelace Respiratory Research Institute
> Albuquerque, New Mexico
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 09:47:40 -0600
> From: bkrummre@ingham.k12.mi.us
> Subject: microscopic anatomy
>
>
> I'm looking for a CD format of microscopic anatomy that is basic for
> Histologic Technicians.
> What I have found in the past is more what is used for medical students
> and diagnostic application.
>
> Any suggestions ??
>
> The intention is for this program to used as a self study curricular
> piece to be combined with hands on/ lab procedure special staining
> techniques.
>
> Betsy Krummrey
> Histologic Technician Program director
> Ingham Intermediate
> 611 Hagadorn Rd
> Mason, Mi 48854
>
>
>
> ******************* NOTE *******************
> There may be important message content
> contained in the following MIME Information.
> ********************************************
>
>
> - ------------------ MIME Information follows ------------------
>
>
> - --------------71A0EE162ECBC7611A29D45B
> Content-Type: text/plain; charset=us-ascii
> Content-Transfer-Encoding: 7bit
>
> <<<<<< See above "Message Body" >>>>>>
>
> - --------------71A0EE162ECBC7611A29D45B
> Content-Type: text/html; charset=us-ascii
> Content-Transfer-Encoding: 7bit
>
> <HTML>
> I'm looking for a <B>CD format of microscopic anatomy </B>that is
<B>basic
> for Histologic Technicians.</B>
> <BR>What I have found in the past is more what is used for medical
students
> and diagnostic application.
>
> <P>Any suggestions ??
>
> <P>The intention is for this program to used as a self study curricular
> piece to be combined with hands on/ lab procedure special staining
techniques.
>
> <P>Betsy Krummrey
> <BR>Histologic Technician Program director
> <BR>Ingham Intermediate
> <BR>611 Hagadorn Rd
> <BR>Mason, Mi 48854</HTML>
>
> - --------------71A0EE162ECBC7611A29D45B--
>
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 09:48:18 -0600
> From: "Jeff Crews"<jcrews@organo.com>
> Subject: Re: prussian blue
>
>      We did this stain recently in bone (which was a real problem due to
>      the decal removing the iron in the marrow). If you have a fine
>      granular staining that overlays the tissue, that's probably
>      precipitate. We had a precipitate problem when we prolonged the
>      staining in bone.
>         What finally worked was filtering the stain initially and at ~15
>      minute intervals with a syringe and Millipore syringe filter. That
>      kept down the accumulation of precipitate.
>      We were doing the stain at 60C in an oven, by the way. And remember
>      that you really shouldn't be seeing a lot of staining in normal
tissue
>      anyway. The RBC's shouldn't be reacting.
>                                                         JC
>
>
> ______________________________ Reply Separator
> _________________________________
> Subject: prussian blue
> Author:  Kathy Walters <kwalters@emiris.iaf.uiowa.edu> at internet
> Date:    11/11/1999 3:16 PM
>
>
>
> Greetings,
>
>         I just finished a Prussian Blue stain run.  I had hardly any
> staining in my liver tissue, light staining in my spleen tissue, and some

> of the staining looked as though it was "over" rather than "in" the
> tissues, and rather punctate.  Does any one still do this stain?  Any
> suggestions?
>
> Kathy Walters                                                    / /
> Research Assistant III                                          / /\
> Center for Microscopy Research                                 / /\ \
> University of Iowa                                            /_/  \ \
> 85 EMRB                                                      ____  ((O))
> Iowa City, Iowa 52242                                        |  |  / /
>                                                               ||  / /
> email:  kwalters@emiris.iaf.uiowa.edu                     -----------
> fax:  (319)335-8049                                      -------------
> www:  http://www.uiowa.edu/~cemrf
>
>
>
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 09:49:20 -0600
> From: "Jeff Crews"<jcrews@organo.com>
> Subject: big honkin' samples
>
>      Does anyone have any experience with embedding and sectioning
>      large-cross-section tissues? We have some porcine vertebral samples
>      that measure about  2" x 1 1/2". I can get larger cassettes for
>      processing them and make some larger embedding molds, but how do you

>      suggest that we clamp them in the microtome? This is the first time
>      that we've had to do something this large, and I'd appreciate
hearing
>      from others about it. Thanks again!
>                                                 jc
>
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 09:49:51 -0600
> From: Karen S Pawlowski <kna101@utdallas.edu>
> Subject: research position available
>
> The Department of Otolaryngology at the University of Texas Southwestern
> Medical Center is currently searching for a person with histology
> experience, preferably parafin experience, to fill a full-time position
in
> our lab.  This person would be trained in the other procedures whenever
> needed.
>
> The medical center is located near downtown Dallas, TX.  For more
> information about this position, please check out the listing for a
> Reasearch Assistant on the ARO (Association for Research in
> Otolaryngology) website:  www.aro.org/positions/other.html
>
> The address for sending resumes is:
>
> Dr. Charles G. Wright
> Dept. of Otolaryngology-Head and Neck Surgery
> UT Southwestern Medical Ctr.
> 5323 Harry Hines Blvd.
> Dallas, TX 75235-9035
>
> If you have any questions, you can contact me via e-mail
> (kna101@utdallas.edu)
>
> Karen Pawlowski
> Sr. Research Assoc./UT Southwestern Med. Ctr.
> PhD candidate/UT Dallas
> Dallas, TX
>
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 10:59:13 -0600
> From: "Jim Staruk" <jstaruk@masshistology.com>
> Subject: Re: big honkin' samples
>
> We've processed specimens like that in the past.  I used an empty plastic
> coverglass container, placed the large specimen in the bottom, placed a
> labeled cassette on top of the specimen and then filled the entire
coverslip
> box with melted paraffin.  After solidification, the specimen was removed
in
> toto and the wax was trimmed so the cassette fit on the chuck.
>
> Jim
> _________________________
> James E. Staruk, HT(ASCP)
> Mass Histology Service
> http://www.masshistology.com
>
>
> - ----- Original Message -----
> From: Jeff Crews <jcrews@organo.com>
> To: <histonet@pathology.swmed.edu>
> Sent: Friday, November 12, 1999 10:36 AM
> Subject: big honkin' samples
>
>
> >      Does anyone have any experience with embedding and sectioning
> >      large-cross-section tissues? We have some porcine vertebral
samples
> >      that measure about  2" x 1 1/2". I can get larger cassettes for
> >      processing them and make some larger embedding molds, but how do
you
> >      suggest that we clamp them in the microtome? This is the first
time
> >      that we've had to do something this large, and I'd appreciate
hearing
> >      from others about it. Thanks again!
> >                                                 jc
> >
> >
> >
>
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 11:00:14 -0600
> From: Ian Montgomery <ian.montgomery@bio.gla.ac.uk>
> Subject: microscopic anatomy
>
> >Date: Fri, 12 Nov 1999 10:16:17 -0500
> >From: Betsy Krummrey <bkrummre@ingham.k12.mi.us>
> >Subject: microscopic anatomy
> >To: Histonet@pathology.swmed.edu
> >
> > I'm looking for a CD format of microscopic anatomy that is basic for
> >Histologic Technicians.
> >What I have found in the past is more what is used for medical students
and
> >diagnostic application.  Any suggestions ??  The intention is for this
> >program to used as a self study curricular piece to be combined with
hands
> >on/ lab procedure special staining techniques.  Betsy Krummrey
> >Histologic Technician Program director
> >Ingham Intermediate
> >611 Hagadorn Rd
> >Mason, Mi 48854
>
> Betsy,
> 	Why not make one yourself. I'm in the process of capturing the
departments
> and my collection of slides then storing the images on CD. Examining
these
> slides again I have found some pretty fabulous fields and with the almost
> limitless capacity of CD's now have a large collection of images. I can
now
> take a set of images, write appropriate text for the course then assemble
> on a CD. The feedback from the students is very, very positive and a
> subject that was always viewed as 'oh no, not ****** histology again' is
> becoming attractive. Only downside, the students don't use a microscope,
> but I can live with that if they now view histology as interesting.
> 	If your worried about cost don't be. All you need is a microscope with
> video camera, Iomega Buz, Adobe photoshop and a CD burner.
> Ian.
>
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 11:45:43 -0600
> From: Linda Prentice <lprent@itsa.ucsf.edu>
> Subject: mouse embryos
>
> Hi
>
> I'm looking for some useful tips on doing paraffin on very late stage
mouse
> embryos (day 18.5) that must be "intact" - meaning that I cannot trim off
> any skin to make fluid exchange easier.
>
> Also, some useful paraffin tips on earlier stage embryos (day 9 and 10)
> would be appreciated.
>
> Thanks
> Linda Prentice
>
> Linda Prentice
> Department of Anatomy
> University of California San Francisco
> 513 Parnassus Ave
> Room S-1343
> San Francisco CA 94143-0452
> Phone 415-476-1439
> Fax 415-476-4845
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 11:46:19 -0600
> From: "Kellar, Eric" <kellarec@MSX.UPMC.EDU>
> Subject: RE: big honkin' samples
>
> Before investing in a Jung Polycut E sliding microtome, I used an old
Leitz
> 820 rotary microtome with an old quick release clamp. I was successful in
> getting pretty nice paraffin whole mount sections on whole larynx and
> prostate specimens. The trick is to get some paraffin support behind the
big
> honkin specimen (approx. 1.5" to 2" of solid paraffin), while still
> achieving enough clearance between the blade and specimen. Also remember
to
> release trapped air bubbles during embedding, if not, they will cause the
> specimen to vibrate and break away from the cassette at the microtome. It
> usually took half a day to properly embed the specimen and then let it
cool.
> Old fashioned lead "L's" worked great as molds with mega-cassettes placed
on
> top.
>
>
>
> Eric C. Kellar
> University of Pittsburgh Medical Center
>
>
>
> > ----------
> > From: 	Jeff Crews[SMTP:jcrews@organo.com]
> > Sent: 	Friday, November 12, 1999 10:36 AM
> > To: 	histonet@pathology.swmed.edu
> > Subject: 	big honkin' samples
> >
> >      Does anyone have any experience with embedding and sectioning
> >      large-cross-section tissues? We have some porcine vertebral
samples
> >      that measure about  2" x 1 1/2". I can get larger cassettes for
> >      processing them and make some larger embedding molds, but how do
you
> >      suggest that we clamp them in the microtome? This is the first
time
> >      that we've had to do something this large, and I'd appreciate
hearing
> >
> >      from others about it. Thanks again!
> >                                                 jc
> >
> >
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 11:47:11 -0600
> From: "Margaret Gondo" <gondom@valentis.com>
> Subject: Re: slidemaster
>
> I heard on the net a few months back that Slidemaster had quit making
> these.  About a year ago, I saw them advertised in a Scytek catalog.
Here
> is the website.  http://www.scytek.com/
>
>
> Good luck
>
>
>
>
>
> Lynn Gardner <lynn-gardner@uiowa.edu> on 11/11/99 12:02:19 PM
>
> To:   histonet@pathology.swmed.edu
> cc:    (bcc: Margaret Gondo/GeneMedicine)
> Subject:
>
>
>
>
> Dear all,
> I am looking for information on manual immuno slide staining trays called
> Slide Masters. We purchased four of them a couple of years ago and I
can't
> find the information and would like to order some more. Could someone
> please send me an e-mail of how to contact the company that sells this
> product.
> As always thanks everyone in advance.
> Lynn Gardner
>
>
>
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 12:38:08 -0600
> From: Gayle Callis <uvsgc@msu.oscs.montana.edu>
> Subject: embedding large sample
>
> Have embedded several ways.
>
> the megacassette can be put on the back of your sample, so the cassette
> sticks up enough (part of the cassette usually clamped in the microtome)
>
> Be sure to have the cassette above the sample, this may require an empty
> cassette spacer below, then add the megacassette, making sure there are
no
> bubbles, and that the megacassette is filled with paraffin, to make
> it more stable.
>
> The old style embedding ring can be applied in the same way, as long as
> you can clamp it into the microtome.  Just make sure you have
> enough of the plastic portion of cassette available to clamp.  A practice
> run maybe?
>
> With that large a bone, I would also process longer, and if anything
> else, do an extra long infiltration with paraffin in the end.
>
> You did not say what kind of microtome you have?  Make sure you have
> the proper clearance for cutting past the knife.
>
> I recall an article on this in the Shandon LabLeader newsletter, you
> could call them and ask for a copy, it had photos to show how the
> embedding ring? or cassette was put on back of a large block.
>
> Gayle Callis
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 13:02:10 -0600
> From: Kevin Fitch <kjf4@cornell.edu>
> Subject: 1150 autocut microtome
>
> Hello everybody,
>
> I am a new subscriber, I am looking for anyone who might have a cassette
> holder for a Reichert-Jung 1150 autocut microtome.  We had this given to
us
> by a retired faculty member at Cornell Univ. He only did plastic
> embedding,  We do wax embedding in my Department. The cassette holder is
no
> longer being manufactured. If anyone could point me into the right
> direction, I would be ever so greatful.
> Also, My Department at Cornell is the Department of Veterinary
Microbiology
> and Immunology. We are a research Laboratory, Our model is Avian Species.
>
> Kevin Fitch kjf4@cornell.edu
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 13:22:59 -0600
> From: "Tim Morken" <timcdc@hotmail.com>
> Subject: Re: big honkin' samples
>
>
> Jeff,
>
> I've made stapeled lenspaper "bags" to process large tissue and "L" irons
to
> make blocks. You can use a 1" cubed wood block or a "histo-ring" plastic
> embedding mount in the paraffin to make a clamp area. You will need to
use
> a screw clamp to hold it. the hard part is making sure the block or
plastic
> ring sits straight in the paraffin as it hardens.
>
> Have fun!
>
> Tim Morken, B.A., EMT(MSA), HTL(ASCP)
> Infectious Disease Pathology
> Centers for Disease Control
> MS-G32
> 1600 Clifton Rd.
> Atlanta, GA 30333
> USA
>
> email: tim9@cdc.gov
>        timcdc@hotmail.com
>
> Phone: (404) 639-3964
> FAX:  (404)639-3043
>
>
> - ----Original Message Follows----
> From: Jeff Crews <jcrews@organo.com>
> To: histonet@pathology.swmed.edu
> Subject: big honkin' samples
> Date: Fri, 12 Nov 1999 10:36:55 -0500
>
>       Does anyone have any experience with embedding and sectioning
>       large-cross-section tissues? We have some porcine vertebral samples
>       that measure about  2" x 1 1/2". I can get larger cassettes for
>       processing them and make some larger embedding molds, but how do
you
>       suggest that we clamp them in the microtome? This is the first time
>       that we've had to do something this large, and I'd appreciate
hearing
>       from others about it. Thanks again!
>                                                  jc
>
>
>
>
>
>
>
>
>
> ______________________________________________________
> Get Your Private, Free Email at http://www.hotmail.com
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 14:15:42 -0600
> From: Amos Brooks <atbrooks@snet.net>
> Subject: Re: Ck7 & Ck20
>
> Hi Dana,
>     I have found quite the opposite true with regards to antigen
retrieval of
> cytokeratins, especially those of high molecular weight. Our prostate
biopsies
> look fantastic with heat induced epitope retrieval (HIER... (I hate
typing all
> that out)).
>     When we optimized this stain, it was suggested that we use an enzyme
> digestion, like Proteinase K (PK). But the stain was much cleaner and
more
> vivid on the HIER tests. Also we noted a greater frequency of tissue fall
offs
> with PK.
>     Each lab is different so it is of utmost importance to play with your
> stains (incubation times, detection kits, antigen retrieval methods etc.)
as
> often as your workflow can allow.
> Amos
>
> DDittus787@aol.com wrote:
>
> > I only have experience with one antibody that you ask about the Ck7,
and it
> > is from Zymed ,it works really well, with a protease digestion and
short
> > incubation time,but i
> > find most cytokeratins work well with digestion rather than retrieval,
and
> > most are affected by oxidation. Hope this helps.       Dana
>
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 17:15:22 -0600
> From: mark.lewis@shandon.com
> Subject: RE: big honkin' samples
>
> Jeff,
>
> Shandon sells a super mega cassette, a super mega cassette base mold  and
a
> quick release clamp to accommodate the super mega cassette block.. It can
be
> used on our microtomes, but am not sure if all microtomes will be able to
use
> these same pieces.  What model of microtome are you using?  Call us if
you
> need more information on these super mega items.
>
> Regards,
>
> Mark Lewis
> Product Specialist
> Shandon
> 1-800-716-3703
>  ----------
> From:  kellarec@MSX.UPMC.EDU [SMTP:MIME :kellarec@MSX.UPMC.EDU]
> Sent:  Friday, November 12, 1999 1:19 PM
> To:  histonet@pathology.swmed.edu; jcrews@organo.com
> Subject:  RE: big honkin' samples
>
> <<File: ENVELOPE.TXT>>
>
--------------------------------------------------------------------------
> --
> Before investing in a Jung Polycut E sliding microtome, I used an old
> Leitz
> 820 rotary microtome with an old quick release clamp. I was successful in
> getting pretty nice paraffin whole mount sections on whole larynx and
> prostate specimens. The trick is to get some paraffin support behind the
> big
> honkin specimen (approx. 1.5" to 2" of solid paraffin), while still
> achieving enough clearance between the blade and specimen. Also remember
> to
> release trapped air bubbles during embedding, if not, they will cause the
> specimen to vibrate and break away from the cassette at the microtome. It
> usually took half a day to properly embed the specimen and then let it
> cool.
> Old fashioned lead "L's" worked great as molds with mega-cassettes placed

>
> on
> top.
>
>
>
>
>
>
> Eric C. Kellar
> University of Pittsburgh Medical Center
>
>
>
>
>
>
> > ----------
> > From:  Jeff Crews[SMTP:jcrews@organo.com]
> > Sent:  Friday, November 12, 1999 10:36 AM
> > To:  histonet@pathology.swmed.edu
> > Subject:  big honkin' samples
> > >      Does anyone have any experience with embedding and sectioning >

>
>     large-cross-section tissues? We have some porcine vertebral samples >

>
>      that measure about  2" x 1 1/2". I can get larger cassettes for >
>
>   processing them and make some larger embedding molds, but how do you >

>
>     suggest that we clamp them in the microtome? This is the first time >

>
>      that we've had to do something this large, and I'd appreciate
hearing
> > >      from others about it. Thanks again!
> >                                                 jc
> > >
>
>
>
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 17:45:30 -0600
> From: "Shotsberger-Gray, Wanda" <WandaShotsberger-Gray@hmhs.com>
> Subject: RE: Need Antigen Retrieval Help
>
> Katie,
> When I HIER, I put all the slides in a 20 slide rack (intended for use on
> the Sakura stainer).  the slides with tissue go in the middle of the
rack,
> and empty slides fill the rest of the slots.  I leave 2 slots in the
middle
> for the microwave temperature probe.  Then the whole works goes into a
> ziplock bag (intended for tissue transport).  I leave a little hole for
the
> probe to go into.
> I "nuke" for 20 minutes at 95 degrees C.  If the stuff boils over, its
> caught in the bag, and the bag makes a little humidity chamber that keeps
> that from happening, somehow.  I used to have trouble with boil over, but
> this cured it.  I have to admit I saw this tip on the histonet a while
back.
> Barry Ritman may hve posted it originaly...
> Wanda Shotsberger
> Harris Methodist
> Fort Worth TX
>  ----------
> From: Bennett, Catherine (Katie)
> To: 'histonet@pathology.swmed.edu'
> Subject: Need Antigen Retrieval Help
> Date: Thursday, November 11, 1999 10:20AM
>
> Hi all!
>
> I'm starting a double staining IHC run this morning that is intending to
> take two days, and I may have already committed a fatal error on step
one,
> so I'm wondering if I should continue or start over.
>
> I am doing microwave antigen retrieval and had a problem with the
solution
> boiling over and completely out of the coplan jar.  My protocol calls for
> microvaving slides in a coplan jar of antigen retrieval for 2x 5 min, but
> after just 1.5 min all the solution is boiled over and gone.  Needless to
> say, I didn't catch it in time and the slides may have gone through some
> microwaving without any solution in the jar.  I had placed the coplan jar
in
> a small dish of shallow water, so the microwave was very humid at least.
>
> I'm guessing the microwave is much more powerful than the one used in the
> protocol I am using and I should experiment with which power setting I
> should use.  However, in the meantime, are the slides from today toast
> because they have been "cooked" without being in the antigen retrieval
> solution, or can I continue with them?
>
> Any suggestions from others on preventing boil-over would also be much
> appreciated.
>
> *********************************
> Catherine "Katie" Bresee Bennett
> Sr. Technical Associate
> Lovelace Respiratory Research Institute
> Albuquerque, New Mexico
>
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 18:00:15 -0600
> From: "P. Emry" <emry@u.washington.edu>
> Subject: penfix vendor?
>
> anyone?
>
>
> Trisha
>
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 18:18:32 -0600
> From: DDittus787@aol.com
> Subject: Re: big honkin' samples
>
> Many Labs do half of Prostate Gland and get a special cassette holder for

> their Microtomes from specialty companies, try Shandon,or Leica, or even
call
> Sakura,the old company Tissua Tek used to carry all that stuff.
>                                              Dana
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 18:19:14 -0600
> From: Katri Tuomala <katri@istar.ca>
> Subject: Re: Ck7 & Ck20
>
> Hi Dana,
> I agree with you that the proteolytic digestion generally works well
> with cytokeratins, but I also agree with Amos on the high molecular
> weight cytokeratin (*clone 34Be12) turning out absolutely best with
> HIER. I've tried pepsin, trypsin and pronase E and there is no
> comparison. On the other hand, cytokeratin Cam 5.2 is most brilliant
> with pronase E. It seems, to get optimal results, each antibody needs to
> be tested with various retrievals or no retrieval. My 2 cents worth...
>
> Katri
>
> Katri Tuomala
> Anatomic Pathology
> St.Joseph's Hospital
> Hamilton, Ontario, Canada
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 19:30:42 -0600
> From: "P. Emry" <emry@u.washington.edu>
> Subject: Re: big honkin' samples
>
> Hi Jeff,
>
> I got a lot of help from others on the net when I started doing large pig
> bones.  I'll pass on what I have learned and used.
>
> I had a fast-release chuck that slipped so that small bites were
> being taken out of the specimens.  I got one of the screw
> down chucks which solved my problems and gave me the ability to use large
> metal bases (aluminum with a grid)  with mega-cassette sized blocks
> attached that fit into the chuck.  I inherited these things with the lab
> so I don't know where you can buy them.
>
> I also use a meg-cassette by filling it with paraffin. It fits nicely in
> the screw down chuck. Unfortunately, the mega-cassette has a hole where
> the hook on the lid fits. When you fill it you have to plug the hole
> with bits of cooling paraffin or it leaks.  I bought a glue gun and
> plugged the hole on a second cassette I use for embedding.
>
> (If there is a vendor listening, redesign the mega-cassette so that hole
> is closed and won't let the paraffin leak out.  Having to use two
> cassettes per specimen is not a good thing and trying to plug it up with
> cooling bits of paraffin is time consuming.)
>
> Am I the only one with this problem or have you real "pros" found a way
to
> avoid this problem?
>
> Best of luck.
>
> Trisha
> U of Washington, Seattle
>
> On Fri, 12 Nov 1999, Jeff Crews wrote:
>
> >      Does anyone have any experience with embedding and sectioning
> >      large-cross-section tissues? We have some porcine vertebral
samples
> >      that measure about  2" x 1 1/2". I can get larger cassettes for
> >      processing them and make some larger embedding molds, but how do
you
> >      suggest that we clamp them in the microtome? This is the first
time
> >      that we've had to do something this large, and I'd appreciate
hearing
> >      from others about it. Thanks again!
> >                                                 jc
> >
> >
> >
>
> Trisha
>
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 21:30:42 -0600
> From: "Noreen S. Gilman" <n4xiu@gate.net>
> Subject: Re: penfix vendor?
>
> Richard Allen Medical
> Noreen
>
> "P. Emry" wrote:
>
> > anyone?
> >
> > Trisha
>
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 21:46:35 -0600
> From: "J park" <jpark100@hotmail.com>
> Subject: digest
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 21:46:59 -0600
> From: "J park" <jpark100@hotmail.com>
> Subject:
>
> digest
>
>
> ----------------------------------------------------------------------
>
> Date: 12 Nov 1999 23:00:45 -0600
> From: Charles Dangler <cdangler@gis.net>
> Subject: Histologist position announcement
>
> Histology Associate
>
> Biogen, Inc., has an immediate opening for a Histology Associate to join
> our expanding team in Cambridge, MA.  Working in a new histology
> laboratory, the selected candidate will provide research support by
> preparing tissues for light microscopic examination and assisting
> scientists by determining the appropriate methods of fixation,
> processing and staining for various research applications.  Primary
> responsibilities will include tissue trimming, processing, microtome and
> cryostat  sectioning, and routine and special stains.  Ability to use,
> maintain and troubleshoot automated laboratory equipment is required.
> Other activities will include ordering and maintaining an inventory of
> supplies and maintaining a histology database.  Requirements: BS or
> Associates degree and demonstrated technical expertise and experience in
> histotechnology.  ASCP certification (HT or HTL) is preferred.  Good
> communication skills, basic computer knowledge, and the ability to work
> well both independently and as part of a team are needed.  Biogen has an
> outstanding record of leadership and accomplishment in the biotechnology
> field, and has a compensation and benefits package, including equity
> participation, that is unmatched in the industry.  Interested candidates
> should FAX their resumes to Human Resources, Source Code TC-HST at (617)
> 679-2546, or Email to resumes@biogen.com (source code only must appear
> on subject line).  Biogen is an Equal Opportunity Employer.  Please
> visit our website at www.biogen.com
>
>
>
>
> Here are the messages received yesterday!