To All,

I have to jump in on this one-I think sometimes we do things because other
techs tell us or else because of first hand knowledge.  I have had occasion
to do both.  When I was employed in a hospital setting speed was very
important (annual case load=80,000).  Sections do curl off a new knife
which tends to be too sharp-soooo---we used to run a wooden stick over the
edge prior to facing or thin sectioning the tissue blocks.  Worked every
time.  As I recall years ago we also used to clean disposable blades before
using them-OIL.  I do not do that now in either job-I cut frozens for a
surgeon and do part time work in paraffin.  As with many other technique
isues in different labs most people do what works for them.  But it is
interesting to see how others do things isn't it??  

Sue Becker, HTL
Albany, NY
----------
> From: HistoNet Server <HistoNet@Pathology.swmed.edu>
> To: HistoNet Server <HistoNet@pathology.swmed.edu>
> Subject: Daily Digest
> Date: Friday, April 09, 1999 3:01 AM
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 01:30:53 -0600
> From: ALLISON@cardiff.ac.uk
> Subject: Re: Mast cell control
> 
> Mast cells in tissue from different species do not all behave the 
> same.  Nor do mast cells at various stages of their own metabolic 
> functions.  This is especially true if you are using metachromatic 
> methods to demonstrate them.
> 
> For Really good demonstration, try alcian blue pH1.0 on frozen 
>  unfixed sections, but do not be surprised if the edges of the cells 
> are a bit fuzzy - unfixed sometimes means "unlocalised" as well.
> Russ Allison, Wales
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 05:45:53 -0600
> From: Ian Montgomery <ian.montgomery@bio.gla.ac.uk>
> Subject: Iomega Buz.
> 
> 	Just bought an Iomega Buz which I installed into a PC then onto a
> video camera on my Zeiss Axiophot. What do I think of the system, superb.
> So far just been capturing images for teaching but now taking images for
> analysis using metamorph.
> 	Does anyone else use the Iomega Buz. If so let me know and we can
> share information.
> Ian.
> 
> Dr. Ian Montgomery,
> West Medical Building,
> University of Glasgow,
> Glasgow,
> G12 8QQ,
> Scotland.
> Tel: 0141 339 8855 Extn. 6602.
> Fax: 0141 330 4100.
> e-mail: ian.montgomery@bio.gla.ac.uk
> 
> 
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 05:58:10 -0600
> From: Decalchem@aol.com
> Subject: Re: cytology preparation techniques book
> 
> 
> In a message dated 4/7/99 11:51:03 PM, jkiernan@julian.uwo.ca writes:
> 
> <<Try:  Boon ME & Drijver JS (1986) Routine Cytological Staining
>         Techniques. Theoretical background and Practice. 
>         New York: Elsevier.
> >>
> 
> This book, and many others for the laboratory references, are available 
> through our website, Http://www.decal-bone.com
> 
> Cliff Berger
> President
> Decal Chemical Corp. 
> email:decalchem@aol.com
> http://www.decal-bone.com
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 05:58:36 -0600
> From: "Hewlett Bryan  (CMH)" <HEWLETT@EXCHANGE1.CMH.ON.CA>
> Subject: RE: Mast cell control
> 
> Dear AEJNSN?
> 
> Any number of tissues can contain mast cells! If one wishes to use them
> as staining controls, one should consider that there two major
> sub-populations of mast cell, which differ considerably in their
> stainability, depending on the staining protocol and the fixative used.
> Connective tissue mast cells are probably the easiest to stain with
> traditional dye technology. Mucosal mast cells can be more problematic,
> particularly with formaldehyde fixation. We use small bowel as a
> control( with a variety of fixatives) because it contains both
> populations, other tissues to consider, for the same reason, would
> include lung and nasal polyp. The number of mast cells demonstrable
> varies with the technology used. In human material, immunohistochemistry
> for CD117 or Mast cell tryptase will reveal the greatest number( in rat
> RMCPI or RMCPII). Dye technology, independent of fixative, will only
> stain approximately 60-90% of mast cells ( paper submitted). I would
> suggest that you read chapter 24, p695, by Lennart Enerbach. In
> "Histochemistry in Pathological Diagnosis". Editor Samuel S. Spicer.
> 1987. Marcel Dekker Inc. NY. ISBN 0-8247-7408-6.
> 
> Hope this helps.
> 
> Bryan
> >----------
> >From: 	AEJNSN@aol.com[SMTP:AEJNSN@aol.com]
> >Sent: 	April 7, 1999 5:16 PM
> >To: 	Histonet@pathology.swmed.edu
> >Subject: 	Mast cell control
> >
> >We are looking for a mast cell control.  In any of the literature I read
it 
> >says any tissue with mast cells in it should work..wel the problem is
finding
> >tissue.  Please do not laugh at such a question but does anyone have any

> >suggestions as to specific tissue that contains mast cells?  Thank you
for 
> >your help.
> >
> >
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 06:21:49 -0600
> From: "Tim Morken" <timcdc@hotmail.com>
> Subject: Re: Mast cell control tissues
> 
> Mast cells are found primarily in the connective tissue of the skin 
> and mucous membranes. They are also found in the thymus and the 
> capsules of most organs. Look around small blood vessels. There are 
> none in the CNS.  The location gives a hint of their function: 
> protection against foriegn elements through quick distribution of 
> chemicals which help eliminate those elements.
> 
> Mast cells are part of the immune system and are reactive to 
> antibodies produced by plasma cells against foriegn elements. The 
> granules in mast cells contain histamine (dialates blood vessels 
> allowing more blood to the affected area) and heparin (prevents blood 
> clotting allowing flushing of tissue), among other things. When you 
> take an anti-histamine you are using a drug that blocks receptors on 
> mast cells and prevents their activation.
> 
> Those who are allergic to bees and other stinging insects have a 
> super-sensitve mast cell system which all release their contents at 
> once causing anaphylaxis.
> 
> 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 
>  
> FAX:  (404)639-3043 
> 
> 
> - ----Original Message Follows----
> From: AEJNSN@aol.com
> To: Histonet@pathology.swmed.edu
> Subject: Mast cell control
> Date: Wed, 07 Apr 1999 18:16:49 -0400 (EDT)
> 
> We are looking for a mast cell control.  In any of the literature I 
> 
> read it 
> says any tissue with mast cells in it should work..wel the problem is 
> 
> finding 
> tissue.  Please do not laugh at such a question but does anyone have 
> 
> any 
> suggestions as to specific tissue that contains mast cells?  Thank you 
> 
> for 
> your help.
> 
> 
> 
> 
>  
> 
> 
> _______________________________________________________________
> Get Free Email and Do More On The Web. Visit http://www.msn.com
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 07:15:42 -0600
> From: rkline@emindustries.com
> Subject: Re: PAS Problems, more info Gayle
> 
> Donna,
> 
> From everything I've read over the histonet, I think all the bases have
> been covered.  I never used the sodium metabisulfite and always had
> excellent results.  I used small intestine as a control for PAS and
> mucicarmine with excellent consistent staining.  I used a flood method
for
> PAS  ( I hated all the dishes).  This would mean the Schiff's would not
be
> sitting in a coplin jar.  My offer to sample the Harleco Schiff still
> stands (no pressure intended).  It may help eliminate the Schiff's as the
> culprit.  I think Schiff's freezing or having ice crystals would be a
> problem.
> 
> Please feel free to call me at 800-222-0342 ext. 443, if you want to
> brainstorm.
> 
> Rande Kline HT (ASCP)
> Technical Services
> EM Science
> 
> 
> 
> 
> Donna Carr <dkc@odsgc.net> on 04/07/99 09:17:05 PM
> 
> To:   'Histonet' <Histonet@Pathology.swmed.edu>
> cc:
> Subject:  Re: PAS Problems, more info Gayle
> 
> 
> 
> 
> Gayle asked for more info.
>     To further elaborate on the problems we are experiencing with our
> PAS stain,  I have worked in Histology for 2 years and never had any
> problem with a PAS stain.  We also use the spit method for diastase.
> The stain has always worked beautifully.  Usually the second the slides
> are placed in the coplin jar the control turns pink.  That's why I am so
> baffled at the recent problems.  The sections are not staining enough.
> They are turning pink but not reacting like they normally do.  We stain
> liver needle biopsies and tumors with our PAS.  We used to reuse our
> reagents but have gone to using a staining rack instead of  coplin
> jars.  I did try the coplin jar method without any success.
> 
> Our method is as follows:
>     hydrate to di H20
>     5 min periodic acid (1g/dl)
>     rinse in DI
>     15 min in Schiff's reagent
>     2 min rinse in Sodium metabisulfate solution
>     2 min rinse in Sodium metabisulfate solution
>     5 to 10 min rinse in running tap water to enhance pink
>     stain 4 min Hematoxylin
>     dehydrate, clear, cover slip
>    this is from memory and I can't remember the concentration of the
> Sodium  metabisulfate?  (0.5g/100ml)
> 
>     1.  We used a different control, so this was the first suspected
> culprit.  We obtained new sections from autopsy liver for controls and
> ran several trials with different old control blocks and new blocks.
> Not seeing any improvement in the staining.
>     2.  We had problems with our reagents having ice crystals in them (
> we share a fridge with chemistry) and have since moved our reagents to a
> different location.  I called both companies that we get our Periodic
> acid and Schiff's reagent from and both assured me that freezing would
> NOT harm to the reagents.  Only to make sure that the Periodic acid was
> thoroughly mixed and at room temp before using.  Concerning the Schiff's
> reagent I was told that it was okay to keep at room temp and no need to
> refrigerate.
>    3. We are a small lab and keep reagents as long as possible.
> 
> My plan of action:
> We have a new bottle of Schiff's that we are now using.  So I think the
> first thing to do is to get a section of cervix to use as a control.
> Secondly to use a new batch of periodic acid.  I don't believe we have
> the chemicals to make our periodic acid, but have a small amount used
> for our fungus stain that is the same concentration.
> 
> Any further suggestions are welcome, sorry so long winded.  Thanks again
> for all the response.
>         Sincerely Donna
> 
> 
> 
> 
> 
> 
> 
> 
> 
> 
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 07:16:13 -0600
> From: rkline@emindustries.com
> Subject: Re: Mast cell control
> 
> Try skin.
> 
> Rande Kline HT (ASCP)
> Technical Services
> EM Science
> 
> 
> 
> 
> 
> AEJNSN@aol.com on 04/07/99 06:16:49 PM
> 
> To:   Histonet@pathology.swmed.edu
> cc:
> Subject:  Mast cell control
> 
> 
> 
> 
> We are looking for a mast cell control.  In any of the literature I read
it
> says any tissue with mast cells in it should work..wel the problem is
> finding
> tissue.  Please do not laugh at such a question but does anyone have any
> suggestions as to specific tissue that contains mast cells?  Thank you
for
> your help.
> 
> 
> 
> 
> 
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 08:00:46 -0600
> From: FreidaC@aol.com
> Subject: Re: PAS Problems, more info Gayle
> 
> Donna:
> 
> This is not too likely an explanation - but since you said that your
Schiff 
> reagent was new- I thought that I might put it out for what it is worth. 
We 
> once got a bottle of Schiff reagent that was intended for use in
Chemistry 
> for the detection of aldehydes, and it gave a much reduced reaction. 
When we 
> got the right Schiff reagent, our problem disappeared.
> 
> Freida Carson
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 08:01:10 -0600
> From: Jamie Erickson <JErickson@genetics.com>
> Subject: Re: Mast cell control
> 
> A good Positive is Skin.....
> 
> Jamie Erickson
> Genetics Institute
> Andover, Ma 
> 
> >>> <AEJNSN@aol.com> 04/07 6:16 PM >>>
> We are looking for a mast cell control.  In any of the literature I read
it 
> says any tissue with mast cells in it should work..wel the problem is
finding 
> tissue.  Please do not laugh at such a question but does anyone have any 
> suggestions as to specific tissue that contains mast cells?  Thank you
for 
> your help.
> 
>                                                                          
    
>                                                                          
    
>                                                                          
    
>                                                                          
    
>                                                                          
    
>                                                                          
    
>                                                                          
    
>                                                                          
    
>                                                                          
    
>                                                                          
    
>                                                                          
    
>                                                                         
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 08:29:05 -0600
> From: FreidaC@aol.com
> Subject: Knives that are too sharp
> 
> I too am surprised by the comments about knives being too sharp.  What 
> criteria are used to determine that a knife is too sharp?  Thanks to all
of 
> you for the responses about cleaning knives before use.  Some of the
answers 
> were surprising.
> 
> Freida Carson
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 08:29:29 -0600
> From: "Williams, Matthew J [PRI]" <MWillia7@prius.jnj.com>
> Subject: exsanguination
> 
> Perhaps I should be more specific as to what it is that I am looking for.
I
> appreciate the responses that I recieved to my first inquiry, but I don't
> think I quite got my point across. The process of anesthetizing and
> exsanguinating lab animals before a necropsy is a procedure that we have
> been doing, at the very least, ever since I started working in this
field.
> It was already a standard procedure then, and it continues to be today.
The
> reasons behind exsanguination are clear enough, also. The tissues fix
more
> thoroughly and we get better sections from tissues such as spleen that
are
> active hematopoietic tissues and as such have a rich blood supply. What I
am
> looking for is documentation, in histo texts, etc. that spell out the
> effects of bleeding out on specimen quality and explain this process so
that
> I can site these sources in support of a discussion on the necessity of
the
> process (can anyone say run-on sentence?). I recieved a copy (thank you,
> Barb Munch) of the 1993 Report of the AVMA Panel on Euthanasia which
clearly
> states that, combined with anesthesia, exsanguination is an acceptable
> method of euthanasia. Nothing that I have been able to find so far has
been
> able to explain the benefit of exsanguination on specimen quality. If
anyone
> out there in Histoland knows where such a reference can be found, I can
be
> reached at 'mwillia7@prius.jnj.com'.
> 
> Thank You.
> 
> Matthew J Williams
> Research Assistant
> Morphologic Pathology
> RW Johnson PRI
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 08:29:59 -0600
> From: FreidaC@aol.com
> Subject: More about PAS
> 
> When I suggested cervix for a PAS control - I intended it for use as a 
> glycogen control.  Kidney is the most sensitive control for the reaction
- as 
> there are fewer reactive sites.  I weakened reaction can be determined
very 
> quickly.  I think I said before on histonet that we stained a multitude
of 
> kidney biopsies - and to  be certain that the results were good - we used
a 
> staining jar of reagent for two batches of slides only.  Whoever poured
the 
> reagent into the jar, could put it back in the refrigerator (and we
always 
> refrigerated).  If you got it from the refrigerator in the staining jar,
then 
> you dumped it after use.  We could always tell when someone tried to
"cheat", 
> because the results showed a decrease in staining.
> 
> As Rande indicated, small intestine is a good control for mucin, but it
is 
> not as sensitive for the quality of the reaction as kidney.  And it goes 
> without saying that a section containing fungi should be used for the PAS

> reaction on tissue suspected of containing fungi.
> 
> Several of the histochemists - Troyer, Vacca, and I think Lillie - state
that 
> because the water supply in many cities is heavily chlorinated and will 
> reoxidize the Schiff reagent if you go directly into running water - it
will 
> increase the possibility of false staining - so I think that the sulfite 
> rinse is probably a good idea.
> 
> Freida
> 
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 09:17:19 -0600
> From: Janice Mahoney <jmahone@nmhs.org>
> Subject: cd-5,cd-23 
> 
> Hello,
> Has anyone had problems working up cd-23, cd-5 or cyclin D1?  We are
> experiencing all sorts of problems.  Advise please.  Vendor, dilution,
> digestion, masking etc., etc.
> Thankyou,
> Janice Mahoney
> Omaha, NE
> 
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 09:17:54 -0600
> From: bamur@alaska.net
> Subject: TB Procedure for Pathology
> 
> Hello to all,
> We are having our CAP next week and have been told that a SOP is needed
> in pathology for TB. If any of you have such would you please fax us a
> copy or let us know where we can get one!
> Our phone # is: (907) 729-1806 and fax is: (907) 729-1807. Hopefully,I
> get several responses!
> Thanks in advance and have a great day and a  good weekend!
> 
> Barbara A. Murray,HT(ASCP)
> The Alaska Native Medical Center
> Anchorage,Alaska
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 10:01:09 -0600
> From: Geoff McAuliffe <mcauliff@UMDNJ.EDU>
> Subject: Re: Mast cell controls and CNS 
> 
> Tim Morken wrote:
> 
> > Mast cells are found primarily in the connective tissue of the skin
> > and mucous membranes. They are also found in the thymus and the
> > capsules of most organs. Look around small blood vessels. There are
> > none in the CNS.
> 
>     I beg to differ. Try Silver et al., 1996, Trends in
> Neuroscience19:25-31; "Mast cells in the brain: evidence and functional
> significance".
>     Also, there are mast cells in the meninges. Dimlich et al. 1991, J.
> Neurocytology 485-503,  "Linear arrays of homogenous mast cells in the
> dura mater of the rat".
> 
> The other comments are right on.
> 
> Geoff
> - --
> **********************************************
> Geoff McAuliffe, Ph.D.
> Neuroscience and Cell Biology
> Robert Wood Johnson Medical School
> 675 Hoes Lane, Piscataway, NJ 08854
> voice: (732)-235-4583; fax: -4029
> mcauliff@umdnj.edu
> **********************************************
> 
> 
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 10:01:32 -0600
> From: Gayle Callis <uvsgc@msu.oscs.montana.edu>
> Subject: mast cells/mucosal in gut or connective tissue
> 
> In general, mast cells will be resident in skin (or connective tissues).
> 
> If you are looking for mucosal mast cells, that is a different story and
> staining technic, also fixative. 
> 
> Gayle Callis
> 
> 
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 10:01:56 -0600
> From: Gayle Callis <uvsgc@msu.oscs.montana.edu>
> Subject: FAS and FasL
> 
> Tried, and then called PharMingen, they did not recommend it for IHC and
it
> was their antibodies.
> 
> Bcl2 works on frozen sections, and there are protocols for paraffin.
> 
> There are TUNEL kits that should work, and really are not that difficult,

> and certainly easier that trying to make IHC work.
> 
> Call Trevigen 800 873-8443
> 
> Gayle Callis
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 10:43:04 -0600
> From: Cynthia Favara <cfavara@niaid.nih.gov>
> Subject: muscle morphology
> 
> All,
> 	I recall methods for getting muscle to eliminate contraction but
> can't put my fingers on the specifics. I recall something about letting
the
> muscle sit on a saline pad for a period of time before fixation. Any
> recollections references etc appreciated. 
> Thanks,
> 
> Cynthia Favara
> Rocky Mountain Laboratories
> 903 S 4th Street
> Hamilton, MT 59840
> ph: 406-363-9317
> FAX: 406-363-9286
> e-mail: cfavara@nih.gov
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 11:15:35 -0600
> From: Gayle Callis <uvsgc@msu.oscs.montana.edu>
> Subject: PAS and Freida's comments
> 
> I think it is also important to know what the pathologists might be
looking
> for, PAS for fungus, glycogen, basement membrane, mucin, etc indicate to
> how I need to do the method.  In fact, when they ask for a PAS for
fungus,
> I use Chromic acid as the oxidizer, not perodic acid.
> 
> Let me count the ways to do PAS!!  I have more than one PAS protocol for 
> different things.
> 
> 
> I also counterstain with hematoxylin for a shorter time, since the
periodic
> acid will enhance hematoxylin staining by converting the DNA/RNA COOH
groups
> to aldehydes, with the chance that you can overwhelm the PAS.  In fact,
this
> is a way to restore hematoxylin staining when tissues have been stored in
> NBF too long, and the pH changes.  A periodic acid step before
hematoxylin
> staining often restores the nuclear staining.
> 
> Full of sidelights today, must be Springtime!
> 
> Gayle Callis
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 11:45:37 -0600
> From: "Barry Rittman" <brittman@mail.db.uth.tmc.edu>
> Subject: Exsanguination
> 
> I cannot recall any references from the literature concerning this. The
> 1993 study mentioned is the only reference of which I am aware.
> The desirability of removal of the blood is however well established.
> For most perfusion techniques the fixative is preceded by a saline or
> buffer wash to remove excess blood and to prevent smaller vesels being
> obstructed by blood being fixed in situ. The major advantage of
> perfusion is not only the rapid delivery of fixative to individual cells
> with a consequent rapid killing time, but the initial hardening of the
> tissue which limits subsequent mechanical damage.
> The major problem with fixation is control of the perfusion pressure and
> opening of vessels which,  under normal circumstances are closed.
> Additionally, some structures such as Schlemm's canal have been reported
> to be decreased in size even when using physiologic intraocular
> pressures.
> Hope this is of use to you.
> Barry
> 
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 11:46:00 -0600
> From: Donna Carr <dkc@odsgc.net>
> Subject: Help, need release form for tissues
> 
> We have a situation in which a patient wants their finger.  But do not
> have a release for such occurrences.  We return gallstones but they are
> rinsed and have no fixative on them. The finger has been in fixative and
> would be released with fixative. Could any of you please fax or email me
> a copy of a release suitable for this situation. (please ASAP)
> It would be greatly appreciated.   Donna
> email:  dkc@odsgc.net
> fax  316-272-2258           Thanks again
> 
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 11:46:29 -0600
> From: rkline@emindustries.com
> Subject: Re: More about PAS
> 
> Freida,
> 
> OPP"s . Thanks for the correction.  I used kidney for demonstrating
> glycogen also.
> 
> If anyone would like to try small intestine for mucin, for PAS reaction
or
> Mucicarmine make sure the tissue has not been sitting in formalin too
long.
> If it's fixed too long the staining is light.  I would get sections at
time
> of autopsy.
> 
> 
> Rande
> 
> 
> 
> 
> 
> FreidaC@aol.com on 04/08/99 10:09:50 AM
> 
> To:   histonet@pathology.swmed.edu
> cc:
> Subject:  More about PAS
> 
> 
> 
> 
> When I suggested cervix for a PAS control - I intended it for use as a
> glycogen control.  Kidney is the most sensitive control for the reaction
-
> as
> there are fewer reactive sites.  I weakened reaction can be determined
very
> quickly.  I think I said before on histonet that we stained a multitude
of
> kidney biopsies - and to  be certain that the results were good - we used
a
> staining jar of reagent for two batches of slides only.  Whoever poured
the
> reagent into the jar, could put it back in the refrigerator (and we
always
> refrigerated).  If you got it from the refrigerator in the staining jar,
> then
> you dumped it after use.  We could always tell when someone tried to
> "cheat",
> because the results showed a decrease in staining.
> 
> As Rande indicated, small intestine is a good control for mucin, but it
is
> not as sensitive for the quality of the reaction as kidney.  And it goes
> without saying that a section containing fungi should be used for the PAS
> reaction on tissue suspected of containing fungi.
> 
> Several of the histochemists - Troyer, Vacca, and I think Lillie - state
> that
> because the water supply in many cities is heavily chlorinated and will
> reoxidize the Schiff reagent if you go directly into running water - it
> will
> increase the possibility of false staining - so I think that the sulfite
> rinse is probably a good idea.
> 
> Freida
> 
> 
> 
> 
> 
> 
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 12:24:22 -0600
> From: "Barry Rittman" <brittman@mail.db.uth.tmc.edu>
> Subject: Prolonged fixation
> 
> A while ago, there was a discussion re how long a formalin fixative for
> EM could be stored before being considered unsuitable. By chance I came
> across an article which addresses this point :
> 
> Mount  SL., Schwarz JE and Taatjes DJ.  Prolonged storage of fixative
> for electron microscopy: effects on tissue preservation for diagnostic
> specimens.
> Ultrastructural Pathology  21:   195-200.  1997. The abstract states:
> 
> "Prolonged storage of Karnovsky's fixative, at both 4 and -20 degrees C,
> is possible in the electron microscope setting. Ultrastructural detail
> was not compromised in specimens processed in fixative that had been
> stored for 6 months. Evidence of smooth muscle and neuroendocrine
> differentiation was present in the form of actin filaments/dense bodies
> and neurosecretory granules, respectively. No difference in preservation
> was detected between specimens fixed in freshly prepared Karnovsky's
> fixative  and fixative that had been stored at either 4 or -20 degrees C
> for up to 6 months. Thuis community hospitals can be provided with
> Karnovsky's fixative on a semiannual basis for surgical pathology
> specimens that may require electron microscopy for diagnosis."
> 
> I guess those of us who prefer to make the fixative up fresh are being
> somewhat conservative.
> 
> 
> Barry
> 
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 12:24:58 -0600
> From: Terri Braud <terri_braud@sfc.sch.org>
> Subject: Release of body parts
> 
> American Myth? or food for thought
> I "heard" of a case where some toes were released to the patient.  The
> patient went home and buried them in his back yard. Several years
> later, the patient moved away and sold the house. The new owners
> decided to put in a flower bed in the back yard and you guessed it, dug
> up the toe bones which led to an extensive police investigation into the
> "discovery of partial human remains".  
> 
> We release identifiable body "parts" to funeral homes only.  :-)   tlb
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 12:53:08 -0600
> From: rkline@emindustries.com
> Subject: Re: PAS and Freida's comments
> 
> The beautiful part about histology, is sometimes there is no right and
> wrong and in accordance with the conversation on using blades and stains,
> variety is the spice of life.  The important part is the end result.
> 
> The other beautiful part is the histonet and it's the never ending
> contributions and opportunities for learning.
> 
> Thanks all,
> 
> Rande
> 
> 
> 
> 
> Gayle Callis <uvsgc@msu.oscs.montana.edu> on 04/08/99 12:40:24 PM
> 
> To:   histonet@pathology.swmed.edu
> cc:
> Subject:  PAS and Freida's comments
> 
> 
> 
> 
> I think it is also important to know what the pathologists might be
looking
> for, PAS for fungus, glycogen, basement membrane, mucin, etc indicate to
> how I need to do the method.  In fact, when they ask for a PAS for
fungus,
> I use Chromic acid as the oxidizer, not perodic acid.
> 
> Let me count the ways to do PAS!!  I have more than one PAS protocol for
> different things.
> 
> 
> I also counterstain with hematoxylin for a shorter time, since the
periodic
> acid will enhance hematoxylin staining by converting the DNA/RNA COOH
> groups
> to aldehydes, with the chance that you can overwhelm the PAS.  In fact,
> this
> is a way to restore hematoxylin staining when tissues have been stored in
> NBF too long, and the pH changes.  A periodic acid step before
hematoxylin
> staining often restores the nuclear staining.
> 
> Full of sidelights today, must be Springtime!
> 
> Gayle Callis
> 
> 
> 
> 
> 
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 12:53:35 -0600
> From: "J. A. Kiernan" <jkiernan@julian.uwo.ca>
> Subject: Re: Mast cell controls and CNS
> 
> On Thu, 8 Apr 1999, Geoff McAuliffe wrote:
> 
> > > capsules of most organs. Look around small blood vessels. There are
> > > none in the CNS.
> > 
> >     I beg to differ. Try Silver et al., 1996, Trends in
> > Neuroscience19:25-31; "Mast cells in the brain: evidence and functional
> > significance".
> 
>  That's true Geoff, though Silver et al were writing about a bird's
>  brain. In rats there are mast cells around some cerebral blood
>  vessels (e.g. Devel. Neurosci. 4:220-224, 1981; Acta Anat. 124:149-158,
>  1985), but perhaps these are not quite truly "in" the brain. 
>  You do get truly intraparenchymal mast cells in parts of the 
>  brains of some mammals, in the habenular nuclei (as in the pigeon) and 
>  also in nearby dorsal and medial parts of the thalamus (e.g. 
>  Nature 210:756-757, 1966; J. Anat. 121:303-311, 1976; Acta Anat.
>  75:443-452, 1970). The only mammals (to my knowledge) that have
>  these truly in-the-brain mast cells, in contact with neurons and
>  glia, are the hedgehog, the tree-shrew and the slow loris.
> 
> > Also, there are mast cells in the meninges. Dimlich et al. 1991, J.
> > Neurocytology 485-503,  "Linear arrays of homogenous mast cells in the
> > dura mater of the rat".
> 
>   The dura mater of the rat has been much used for experiments
>   with mast cells. See big book, "The Mast Cells" by Hans Selye
>   (Butterworths, 1963). This would be a good +ve control tissue;
>   so would rat or mouse external ear or tongue, which also
>   contain lots of connective tissue-type mast cells. Rodent
>   mast cell granules survive pretty well any fixation. For other
>   species an alcoholic fixative will give the best results
>   (Carnoy and alcoholic Bouin are both OK).
> 
>  John A. Kiernan,
>  Department of Anatomy & Cell Biology,
>  The University of Western Ontario,
>  LONDON,  Canada  N6A 5C1
> 
>  
> 
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 12:54:06 -0600
> From: rkline@emindustries.com
> Subject: Re: Release of body parts
> 
> Terri,
> 
> We had a case where we had to release a patient's leg to his family for
> burial.  It was for religious reasons.
> 
> Rande
> 
> 
> 
> 
> Terri Braud <terri_braud@sfc.sch.org> on 04/08/99 02:09:15 PM
> 
> To:   histonet@pathology.swmed.edu
> cc:
> Subject:  Release of body parts
> 
> 
> 
> 
> American Myth? or food for thought
> I "heard" of a case where some toes were released to the patient.  The
> patient went home and buried them in his back yard. Several years
> later, the patient moved away and sold the house. The new owners
> decided to put in a flower bed in the back yard and you guessed it, dug
> up the toe bones which led to an extensive police investigation into the
> "discovery of partial human remains".
> 
> We release identifiable body "parts" to funeral homes only.  :-)   tlb
> 
> 
> 
> 
> 
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 14:15:27 -0600
> From: "Michael J. Lyon, Ph.D." <lyonm@VAX.CS.HSCSYR.EDU>
> Subject: Factor VIII or von Willebrand Factor
> 
> Since the original question came on the Histonet I also have a question
to
> those that have used this antibody.  While my efforts have been
successful,
> the resulting fluorescence is blotchy within the endothelial cells.  I
was
> wondering if this is the normal appearance or have I done something
wrong.
> We would like to use this stain to identify the vascular elements within
a
> section but also to identify and innervation.  We are using confocal
> microscopy so the blotchy staining give discontinuous images.
> 
> Any suggestions welcome.
> 
> Mike
> 
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 15:46:10 -0600
> From: "Barnhart, Tammy" <tbarnhart@primecare.org>
> Subject: RE: release of body parts
> 
> We release all kinds of body parts.  Placentas (Native Americans will
wear a
> small portion of it in a bag around their necks.  Good luck for the
child),
> toes, fingers and limbs.  North Dakota has lax burial laws.  You can bury
> anything in your back yard.  We had a woman who wanted her toes so she
could
> bury them with her left leg (lost in a farming accident), four fingers
> (same) and three fetuses she had lost.   Needless to say I have a rather
> complicated specimen release form and system.  You just never know......
> 
> Tammy Barnhart, BS,HTL(ASCP)
> Anatomic Pathology Supervisor
> Pathology Consultants/St. Alexius Medical Center
> Bismarck, ND
> tbarnhart@primecare.org
> 
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 15:58:31 -0600
> From: "John Shaw" <jolesh@hotmail.com>
> Subject: Mast Cells and Copper Controls
> 
> I have found that your local vet can be a good source of control 
> tissues.  For copper, dog liver is an excellent control.  For mast 
> cells, a cross-section of rat ear has wonderful mast cells lined-up 
> along either side of the cartilage.
> 
> 
> _______________________________________________________________
> Get Free Email and Do More On The Web. Visit http://www.msn.com
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 16:46:04 -0600
> From: Cindy Farman <cfarman@sierrabiomedical.com>
> Subject: RE: Mast Cells and Copper Controls
> 
> Dog liver is only a good control for copper if it contains excessive 
> quantities of copper, an abnormal situation that is not very common. So
if 
> you just were to stain regular dog liver for copper you will not see any.
> 
> Cindy Farman
> 
> - -----Original Message-----
> From:	John Shaw [SMTP:jolesh@hotmail.com]
> Sent:	Thursday, April 08, 1999 2:54 PM
> To:	HistoNet@Pathology.swmed.edu
> Subject:	Mast Cells and Copper Controls
> 
> I have found that your local vet can be a good source of control
> tissues.  For copper, dog liver is an excellent control.  For mast
> cells, a cross-section of rat ear has wonderful mast cells lined-up
> along either side of the cartilage.
> 
> 
> _______________________________________________________________
> Get Free Email and Do More On The Web. Visit http://www.msn.com
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 17:16:04 -0600
> From: "Jim Manavis" <jim.manavis@imvs.sa.gov.au>
> Subject: re:cd-5
> 
> On our FFPE material we use a CD-5 from Becton Dickinson (clone leu-1) at
a
> dilution of 1:10 with TRS microwave retrieval, and obtain very good
results.
> 
> As far as Cyclin D1 is concerned we use the Novocastra (NCL-CYCLIN D1-GM)
> antibody at a dilution of 1:60 with TRS microwave retrieval, Trypsin, and
> enhancement with Imidazole. Works beautifully.
> 
> Jim Manavis
> Neuropathology & ICC Lab
> Dpt Tissue Pathology
> IMVS
> 
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 18:01:06 -0600
> From: TABrecken@aol.com
> Subject: Job Postings
> 
> The Florida Society for Hsutotechnology will be displaying a job posting 
> board at our annual seminar April 30-May 2, 1999.  Anyone interested in 
> posting available positions, please email TABrecken@aol.com with the 
> information.
> Thanks.
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 19:00:11 -0600
> From: MFAIRM@aol.com
> Subject: Hales
> 
> Does anyone happen to have a procedure for the Hales Colloidal Iron
stain?  
> We don't have it in any of our (few) reference books.
> 
> Mara Fairman
> Roper Hospital 
> Charleston, SC
> Fax: 843-724-2356
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 19:30:20 -0600
> From: Elizaha@aol.com
> Subject: Source for Lewis antibodys
> 
> Hi,
>  Does anyone have a source for  Lewis antibodies: Lewis A, B, X,Y and 
> sialyl-Le X  
>   (the whole panel)
> 
>     Thanks,
>     Elizabeth
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 20:15:17 -0600
> From: Janice M Pfluke <jpfluke1@juno.com>
> Subject: CEA
> 
> Fellow Histonetters..Can any of you help me with the address and phone
> number for either Hybritech or Immunotech. I am looking for antibody CEA
> (clones CEJ065 and A5B7) and have been told these companies carry one or
> the other, but I do not currently have a catologue for either one.
> Thanks!
> 
> Jan Pfluke
> Histology/IMH
> Rochester General Hospital
> Rochester, N.Y.
> 
> ___________________________________________________________________
> You don't need to buy Internet access to use free Internet e-mail.
> Get completely free e-mail from Juno at http://www.juno.com/getjuno.html
> or call Juno at (800) 654-JUNO [654-5866]
> 
> 
> ----------------------------------------------------------------------
> 
> Date: 8 Apr 1999 22:15:26 -0600
> From: fayerae@digisys.net
> Subject: Temp positions
> 
> I was wondering about the traveling tech services and what have been
> other peoples experience with them. Has your hospital employed any and
> which service did you use. Has anyone out there been a traveling tech
> and how did you like it?
> This isn't a technical question but hope some of you wouldn't mind
> helping me out  with my curiosity.
> Much Thanks
> Jennifer
> 
> 
> 
> Here are the messages received yesterday!