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-----Original Message-----
From: HistoNet Server <HistoNet@Pathology.swmed.edu>
To: HistoNet Server <HistoNet@pathology.swmed.edu>
Date: Sunday, November 22, 1998 5:26 PM
Subject: Daily Digest


>
>----------------------------------------------------------------------
>
>Date: 20 Nov 1998 05:31:15 -0600
>From: "Hessling, Rene" <Hessling@biologie.uni-osnabrueck.de>
>Subject: Neural markers for annelids
>
>Dear Histonetters!
>
>We are studying the structure and organization of nervous systems in
>polychaetes (segmented and mostly marine "worms") and other annelids in
>a phylogenetic context. We label our specimens using antibodies against
>various neurotransmitters or acetylated alpha-tubulin e.g. Unfortunately
>staining against tubulin does not seem to label neurotubuli in the
>species I am currently working on, even though this antibody has proven
>to work perfectly in many other species.
>
>Therefore I am looking for alternative antibodies in order to stain
>neural pathways. Antibodies against neurofilaments would probably be
>suitable, yet suppliers generally offer ones which are only against
>vertebrate tissue. I would appreciate any help or suggestions.
>
>Thanks in advance.
>
>Rene
>
>
>........................................................................
>.............................
>Dipl. Biol. Rene Hessling
>
>Universitat Osnabruck
>FB Biologie/Chemie
>Spezielle Zoologie
>Barbarastr. 11
>49069 Osnabruck
>GERMANY
>........................................................................
>.............................
>
>
>
>
>
>
>
>
>----------------------------------------------------------------------
>
>Date: 20 Nov 1998 08:38:45 -0600
>From: andrea_kelly@CCGATEWAY.AMC.EDU
>Subject: Re:
>
>     You can get them from Wasatch Histo Consultants. Cathy Mayton-(702)
>     625-4425 phone/fax or histology@the-onramp.net
>
>     Andrea Kelly
>     Albany Medical College
>
>
>______________________________ Reply Separator
>_________________________________
>Subject:
>Author:  Janet Golden <JGolden@genetics.com> at Internet-Mail
>Date:    11/19/98 4:59 PM
>
>
>Does anyone use white or opaque glass or
>plastic slides, 1" x 3" ?  If so, please give
>some info as to where and from whom I can
>order them from.
>
>Thanks
>
>
>
>
>----------------------------------------------------------------------
>
>Date: 20 Nov 1998 09:15:06 -0600
>From: Cheryl Crowder <crowder@vt8200.vetmed.lsu.edu>
>Subject: Luxol Fast Blue
>
>I have not been able to access the Histonet on "Send".  This message has
>been sent before and I am trying again.
>
>
>Lauren - There are several factors that you need to consider before
becoming
>"paranoid" about the LFB.  Since alcohol has a lower boiling point than
>water, it will boil faster than water.  Therefore, when heated with
>microwaves, the solution heats faster.  Since many techs heat all solutions
>at the same power setting and time, the solution may boil and mess up the
>cavity of the MW.
>        First, you should heat your alcohol, say 30 seconds, stir the
>solution with a thermometer and read the temperature.  If it is 55 degrees
>C, then you know that the time and power setting for that alcohol-based
>stain.  If the temperature is lower than 55, add 5 more seconds with room
>temperature alcohol and repeat the procedure.  Obviousely, if the temp is
>over 55C, you need to reduce the power level or time.  If you keep the
>alcohol at or below 55C, there is no problem with boiling or sparks.
>        To contain the fumes of the alcohol, I suggest you put the
container
>in a freezer-weight or better zip-type bag.  The solution is then heated,
>the bag is taken to a properly ventilated area, and opened.
>        There should be no problems at all with the alcohol-based stain in
>the MW if you take a few precautions and do come QC on the oven.
>
>Cheryl Crowder
>LSU - School of Veterinary Medicine
>
>
>
>
>----------------------------------------------------------------------
>
>Date: 20 Nov 1998 10:31:12 -0600
>From: Heather McKinnon <heather.mckinnon@pharmagene.com>
>Subject: Chicken antibodies
>
>
> Hi,
>
> Can anyone give me some help about chicken antibodies? I've just
>bought a primary antibody raised in chickens which is IgY fraction (Aves
>Labs). They said that Vector have an anti-chicken IgY secondary, but
>Vector only have an anti-chicken IgG and I don't know if these are the
>same. I read somewhere that IgY is the avian form of mammalian IgG but I
>don't know if this is true. Can anyone help??
>
> Thanks a lot,
>
> Heather.
>
>
>----------------------------------------------------------------------
>
>Date: 20 Nov 1998 10:32:33 -0600
>From: "J. A. Kiernan" <jkiernan@julian.uwo.ca>
>Subject: Re: myelin stain
>
>On Thu, 19 Nov 1998, Karen D. Larison wrote:
>
>> A graduate student here is attempting to stain 50 micron brain sections
for
>> myelin using the Gallyas stain, but isn't having much luck with stain
>> penetration.  Is there anything he can add to his solutions to increase
>stain
>> penetration, or is there an alternate myelin stain he could use on these
>thick
>> sections?
>
>   Gallyas has devised several silver techniques, for a wide range
>   of uses. They are what Manfred Gabe (1976 book) would have called
>   "difficult" methods - meaning ones with several steps that go
>   wrong if you make any sort of mistake. This includes the mistake of
>   not knowing the reasons for all the steps! Why not use a simple
>   method for a simple job like staining myelin in frozen sections.
>
>   The easiest is to use a fat-soluble dye. Sudan black B does a
>   great job, but is likely to make sections as thick as 50 um
>   black all over, because there's myelin everywhere. If the reason
>   for the staining is anatomical orientation - distinguishing
>   white matter from grey - then one of the less strongly coloured
>   Sudan dyes (Sudan III or IV or oil red O) would be more
>   suitable. A slower but also easy stain is luxol fast blue MBS.
>   This works well with thickish frozen sections; you differentiate
>   until the colour as as you like it. All the histological techniques
>   books have these simple methods. If this graduate student is hoping
>   to look for features of individual myelinated nerve fibres, then
>   much thinner sections will be needed and a larger repertoire of
>   techniques is available, depending on the nature of the enquiry.
>   The student's supervisor ought to know all this and be giving advice
>   derived from superior knowledge and longer experience :-(
>
>   Hope this helps.
>
> John A. Kiernan,
> Department of Anatomy & Cell Biology,
> The University of Western Ontario,
> LONDON,  Canada  N6A 5C1
>
>
>
>
>
>
>----------------------------------------------------------------------
>
>Date: 20 Nov 1998 10:33:03 -0600
>From: "Patricia M. Karlisch" <pkarlisch@psghs.edu>
>Subject: Daily Digest -Reply
>
>Histonetters,
>I apologise - the heading for the HT position should read Pa/HT
(Pennsylvania
>HT)  not PA/HT -Pathology Assistant-HT.  Thanks
>PKarlisch
>GMC
>Danville, PA
>
>
>
>----------------------------------------------------------------------
>
>Date: 20 Nov 1998 13:27:53 -0600
>From: Gayle Callis <uvsgc@msu.oscs.montana.edu>
>Subject: chicken antibodies
>
>Jackson ImmunoResearch 800 367-5296, they have a donkey antichicken
>IgY (IgG in parenthesis) H+L, absorbed to multiple species, purified
>and conjugated to biotin, HRP, AP, fluorochromes.  Prices are excellent.
>
>Others Kirkegaard Perry
>Rockland Immunological
>Biodesign
>Harlan aka Serotek, may also have what you need.
>
>Good luck,
>Gayle Callis
>
>
>----------------------------------------------------------------------
>
>Date: 20 Nov 1998 13:28:36 -0600
>From: Nita Searcy 949-6843 <SEARNJ@Integris-Health.com>
>Subject: problem
>
>Help! B5 fixed bone marrows, placed on + slides, left overnite in low temp
>oven, Dako Immunostainer, SECTIONS are washing! No problems with any other
>stains ran at same time . Any ideas? Thanks
>
>
>
>----------------------------------------------------------------------
>
>Date: 20 Nov 1998 13:28:59 -0600
>From: Linda Jenkins <jlinda@ces.clemson.edu>
>Subject: Slides
>
>Janet:  Try Wasatch Histo Consultants at 702.625.4425. That's where I get
>my white plastic slides.
>
>*********************************
>Linda Jenkins, HT
>Clemson University
>Department of Bioengineering
>Clemson, SC
>**********************************
>
>
>
>
>----------------------------------------------------------------------
>
>Date: 20 Nov 1998 16:20:13 -0600
>From: "Kellar, Eric" <kellarec@MSX.UPMC.EDU>
>Subject: RE: myelin stain
>
>Karen:
>
>As John Kiernan has mentioned, these methods by Gallyas* are difficult.
They
>are especially tricky to trouble-shoot on such thick sections. A
>trouble-shooting chart is available on below reference.
>
>If the student is attempting the Gallyas technique for neurofibrillary
>changes in CNS, then the preparation of the stock solutions for the working
>developer is critical. The order of the chemicals into the DH20, must be
>dissolved thoroughly and consecutively.
>
>If another technique is an option, I would also suggest the Luxol fast
>blue/cresyl violet or the Luxol fast blue/PAS method (Kluver and Barbera).
>
>*Gallyas, F. , Hsu, M. , and Buzsaki, G. Four modified silver methods for
>thick sections of formaldehyde-fixed
>     mammalian central nervous tissue: 'dark' neurons, perikarya of all
>neurons, microglial cells and capillaries. J. Neurosci.
>     Methods 50:159-164, 1993.
>
>Eric Kellar
>Histology/Immunohistochemistry
>University of Pittsburgh Medical Center.
> ----------
> From:  Karen D. Larison [SMTP:LARISONK@UONEURO.uoregon.edu]
> Sent:  Thursday, November 19, 1998 10:06 PM
> To:  HistoNet@Pathology.swmed.edu
> Subject:  myelin stain
>
> Histonetters,
>
> A graduate student here is attempting to stain 50 micron brain
>sections for
> myelin using the Gallyas stain, but isn't having much luck with
>stain
> penetration.  Is there anything he can add to his solutions to
>increase stain
> penetration, or is there an alternate myelin stain he could use on
>these thick
> sections?
>
> I am so greatful to this community of histologists for their
>knowledge and
> their willingness to share their expertise.
>
> Karen Larison
>
>
>
>
>----------------------------------------------------------------------
>
>Date: 20 Nov 1998 16:21:22 -0600
>From: "Woodfin, Amy C" <AWoodfin@peacehealth.org>
>Subject: Slides out by 0800...
>
>Dear Histonetters:
>
>We seem to continuously be looking at ways to get our slides out earlier
>each day.  Not a bad thing, just a little daunting when processing times,
>staffing, and equipment limitations are taken into consideration.  Not that
>anyone else ever has this problem...right?
>
>We are starting a formal Quality Improvement process to look at our current
>TAT for cases; more specifically the time all of the slides are out to the
>pathologists.  I/we would be GREATLY appreciative of input on the following
>questions.  Feel free to respond by e-mail, print a copy and fax
>it...however is easiest for you.
>
>Thank you in advance for taking time out of your day to help us with this
>information.
>
>Amy C. Woodfin
>Pathology Supervisor
>St. Joseph Hospital
>2901 Squalicum Pkwy
>Bellingham, WA  98225
>(360) 738-6340
>(360) 738-6776 FAX
>
>***************************************************************************
*
>**********
>SURVEY:
>
>Size of Your Facility:
># of Other Hospitals/Labs within a 90 Mile Radius:
>
>Avg # Cases/Day:
>Avg # Blocks/Day:
>
># of Pathologists:
># of Histotechs:
>Shifts:
>(example:  1 @0400 8hrs, 1 @0600 8hrs, 1@ 0630 8hrs)
># of Microtomes:
>
># of Processors:
>Types of Tissue in each:
>Length of Cycle and End Times:
>
>Routine Staining Process:  Automated?  Manual?
>Special Stain Process:  Automated?  Manual?
>Coverslipping:  Automated?  Manual?
>
>Normal Time Slides out to Pathologist(s):
>Average Case TAT:
>
>May I contact you directly with further questions?
>
>Thanks again!
>
>
>----------------------------------------------------------------------
>
>Date: 20 Nov 1998 16:21:47 -0600
>From: Carla_Aiwohi@usgs.gov (Carla Aiwohi)
>Subject: Spurr's thanks
>
>Thanks to all who responded to my question about rapid cure vs. hard Spurr
>embedding mixtures.  Your info really helped!
>
>Thanks again,
>
>Carla Aiwohi
>
>
>----------------------------------------------------------------------
>
>Date: 20 Nov 1998 16:22:11 -0600
>From: maria@skivs.ski.org (Maria Mejia)
>Subject: Antibody search
>
>Hello everyone. I'm looking for an antibody specific for retinal
>ganglion cells. Does anyone have a source for this antibody?
>Any info you can give me is most appreciated.
>Also on another topic does anyone out there have information
>on designing and building a new workable research histology lab?
>Again thanks for any information you can give me.
>all the best
>Maria Mejia
>Smith-Kettlewell Eye Res. Inst.
>S.F. Ca.
>email address maria@skivs.ski.org
>
>
>----------------------------------------------------------------------
>
>Date: 20 Nov 1998 18:28:36 -0600
>From: Shirley Powell <powell.s@gain.mercer.edu>
>Subject: Re: problem
>
>In the past I too have had problems with B5 fixed tissues washing off the
>positive charged slides.  I investigated and found that the tissues that
were
>post fixed in B5 following Formalin fixation were the ones that washed the
>most.  It was not a constant happening but I changed my protocol and used
>StayOn from Surgipath in my water bath for all tissues that were post B5
fixed
>and used regular slides.  I do not know the chemistry but it keeps the
>sections on the slides run on my NexES.  Maybe the chemical gurus can
explain
>the why, maybe it changes the charge or something.
>
>Shirley Powell
>
>Nita Searcy 949-6843 wrote:
>
>> Help! B5 fixed bone marrows, placed on + slides, left overnite in low
temp
>> oven, Dako Immunostainer, SECTIONS are washing! No problems with any
other
>> stains ran at same time . Any ideas? Thanks
>
>
>
>
>
>----------------------------------------------------------------------
>
>Date: 20 Nov 1998 18:28:59 -0600
>From: andreah@imclone.com
>Subject: Histo workshops
>
>Does anyone know how I could find out if and when and where there
>are histology/histotechnology/pathology etc etc workshops or courses in
>the new york area?  Any suggestions would help ...
>
>Andrea Hooper
>ANDREAH@IMCLONE.COM
>
>
>
>
>
>----------------------------------------------------------------------
>
>Date: 21 Nov 1998 10:30:29 -0600
>From: rueggp@earthlink.net
>Subject: tissue hazards
>
>folks,
>i am now working with rabbit joints that have been injected with snake
>venom.  what are the possible hazards of working with such samples.
>they are fixed in 10% NBF as whole limbs, i cut them into and fix
>further before decalcifing.  i of course use gloves to handle them.  i
>am a super allergic person to venom (bee venom).  should i take further
>percautions.
>patsy ruegg
>
>
>
>----------------------------------------------------------------------
>
>Date: 21 Nov 1998 10:31:46 -0600
>From: "tylee" <tylee@itis.com>
>Subject: Re: Chicken antibodies
>
>Heather,
>
>If it is anti-chicken, it probably means it will recognize the IgY you have
>ordered. The anti-chicken IgG may have been named that way either: 1) for a
>marketing reason to sell to those not familiar with IgY or 2) because the
>technical writer was not familiar with the IgY notation used for chickens.
>I am not an expert; but, I do not think chickens produce IgG. For a brief
>explaination I saw a few years ago with a few references, you can see
>http://www.promega.com/pnotes/46/2259e/2259e.html.
>
>Ty Lee
>- ----------
>From: Heather McKinnon <heather.mckinnon@pharmagene.com>
>To: 'HistoNet@Pathology.swmed.edu'
>Subject: Chicken antibodies
>Date: Friday, November 20, 1998 8:40 AM
>
>
> Hi,
>
> Can anyone give me some help about chicken antibodies? I've just
>bought a primary antibody raised in chickens which is IgY fraction (Aves
>Labs). They said that Vector have an anti-chicken IgY secondary, but
>Vector only have an anti-chicken IgG and I don't know if these are the
>same. I read somewhere that IgY is the avian form of mammalian IgG but I
>don't know if this is true. Can anyone help??
>
> Thanks a lot,
>
> Heather.
>
>
>
>----------------------------------------------------------------------
>
>Date: 21 Nov 1998 10:32:50 -0600
>From: Masayuki Miyagishima <mmiyagis+@pitt.edu>
>Subject: NF-kB immunohistochemistry
>
>Again, I would like to seek some help from histonet.
>I want to show the nuclear localization of activated NF-kB in the heart
>of trangenic mice (myocarditis model).  Any informatin related to NF-kB
>immunohistochemistry is welcome.
>Thank you very much in advance.
>
>Masayuki Miyagishima, MD
>University of Pittsburgh,
>Department of Surgery
>
>
>----------------------------------------------------------------------
>
>Date: 21 Nov 1998 10:45:55 -0600
>From: poobear31@webtv.net (K M)
>Subject: Employment Opportunities
>
>Hi everyone.  I'm new on your email loop so any help you can provide
>would be appreciated.  I was wondering if any of you know of any
>positions available in the tri-state area of De/Pa/Md.  I have an
>associates degree in histotechnology, but unfortunately have not been
>able to find employment in a histo lab.  Currently I'm employed in a
>private lab & have been cross trained in the general lab area.  I love
>my job but would like to get into histo before I've been out too long to
>get back in.  Any comments or suggestions would greatly be appreciated.
>Thank you,
>Kim <aka>  poobear31@webtv.net
>
>
>
>----------------------------------------------------------------------
>
>Date: 21 Nov 1998 10:46:23 -0600
>From: "Jeff Silverman" <peptolab@hamptons.com>
>Subject: Billing for recuts,vs send outs.
>
>Clinical Netters-
>What are your labs doing, charge wise,  when the patient picks up slides
>for referral to a treating institution- either recuts or sending originals?
> Is there a charge to the patient for picking up either recuts or original?
> Do you charge for copies of reports only, clerical retrieval, review by
>docs, or only if you have to reproduce the slides by recuts, or never? Hope
>this isn't too confusing, but I'd like to know.
>
>Jeff Silverman
>peptolab@hamptons.com
>
>
>Here are the messages received yesterday!
>