RE: Daily Digest

From:DRitter@stmarygj.com

Hello
Does anyone have a great T cell marker that they are using for
Immunotperoxidase technique? We are having a few problems with the CD-3
marker we are currently using like to much overall staining making
differentiation impossible.
Any sugestions would be aprreciated. We do have adequate blockings and
just can't get any consistent results.

Thanks

Debra Ritter
Histology dept
St. Marys Hosp.
Grand Junction, Co.
970-244-2080

>----------
>From: 	HistoNet Server[SMTP:histonet@pathology.swmed.edu]
>Sent: 	Tuesday, April 03, 2001 1:00 AM
>To: 	HistoNet Server
>Subject: 	Daily Digest
>
>
>----------------------------------------------------------------------
>
>Date: 2 Apr 2001 01:41:11 -0600
>From: ALLISON@CARDIFF.AC.UK
>Subject: Re: Gold taning 
>
>John,
>
>Is that an "April fool" response?
>Try reciting the authors' names out loud!
>
>
>Russ Allison, 
>Dental School
>Cardiff
>Wales
>
>
>----------------------------------------------------------------------
>
>Date: 2 Apr 2001 06:51:36 -0600
>From: "Krahn, Daniel" <DKrahn@nmh.org>
>Subject: Competency assessment 
>
>Hello all,
>	First off, I apologize for posting another request for competency
>testing info, but I would really appreciate any information that you could
>share with me on competency testing in your departments?  For example, what
>is your process, and what skills are you observing for the individual
>positions?  Any example forms that you could share?
>
>Thank you very much,
>Daniel Krahn
>
>
>----------------------------------------------------------------------
>
>Date: 2 Apr 2001 09:30:46 -0600
>From: Darren Robinson <drobinson@ciit.org>
>Subject: Polycut microtomes
>
>I have a microtome question for all you big specimen sectioners out 
>there.  Has anybody ever used the Leica Polycut-S in a freezer before, and 
>if so did you need to change the set up, i.e. did you replace the 
>lubricating oils to account for the low temperatures.  I am thinking about 
>trying to cut some large frozen specimens and would like to know whether or 
>not I'm about to break an expensive piece of machinery.
>
>****************************************
>Darren Robinson
>Postdoctoral Fellow
>CIIT Centers for Health Research
>6 Davis drive
>Research Triangle Park
>North Carolina
>27709
>
>Tel. 919-5581385
>Email. DRobinson@ciit.org
>****************************************
>
>
>
>----------------------------------------------------------------------
>
>Date: 2 Apr 2001 10:05:25 -0600
>From: Toosandyd@aol.com
>Subject: NY State Meeting May 4-5th
>
>Reminder: Hotel reservations for the meeting need to be in by April 13th. You
>can make your reservations by calling the Wyndham at 315-432-0200. Rate is 
>$99.00.
>
>If you did not receive a program, please let me know I will will get one out 
>to you.
>
>Sandy :)
>
>
>----------------------------------------------------------------------
>
>Date: 2 Apr 2001 11:12:02 -0600
>From: lbrennan@coh.org
>Subject: Thermanox Plastic  Coverslips
>
>We use Thermanox plastic coverslips that cultures have been growing on for
>a week.  We  fix, stain, dehydrate through acetone and  histoclear.   The
>only problem we have had is not dehydrating long enough because the tissue
>is 250 um thick.
>
>     Lynn Brennan
>     City of Hope med center
>     Duarte, CA
>
>
>
>----------------------------------------------------------------------
>
>Date: 2 Apr 2001 11:31:05 -0600
>From: Patsy.Ruegg@UCHSC.edu
>Subject: IHC References
>
>IHC Resource Group Reference List        UPDATED 5-99 PR
>
>In review by the IHCRG  03-01
>Dr Shi and Clive Taylor's book on Antigen Retriveal 
>
>Jules Elias, Immunohistopathology, A practical approach to Diagnostics,
>ASCP Press, ISBN # 0-89189-300-8
>
>Immunomicroscopy: A Diagnostic Tool for the Surgical Pathologist. V19.
>Taylor, Clive.  W.B Saunders Company.  Philadelphia, PA  1986.
>
>Applied Immunohistochemistry for the Surgical Pathologist.  Anthony S-Y
>Leong  Edward Arnold, 1993
>
>In Situ Hybridization.  Royal Microscopical Society Microscopy Handbooks.
>Leitch, A.R et al.  Bios Scientific Publishers Ltd.  1994.
>
>Advances in Immunohistochemistry. DeLeihs, Ronald A. Raven Press 1998
>
>Atlas of Diagnostic Immunohistopathology.  True, Lawrence D.  J.B.
>Lippincott Co.  1990
>
>Handbook of Laboratory Solutions.  By MH Gabb and WE Latchem:  Chemical
>Publishing Company, Inc.  Ny, NY.
>
>Principles of Immunodiagnostics:  Aloisi RM:  The C.V.   Mosby Company;
>1979
>
>Textbook of Immunology;  Barrett JT:                The C.V. Mosby Company,
>1988
>
>Principles and Techniques in Diagnostic Histopathology,  Developments in
>Immunohistochemistry and Enzyme Histochemistry;  Elias J:  
>Noyes Publications, 1982
>
>PCR Technology.  Principles and Applications for DNA Amplification;
>Erlich HA (editor):  M. Stockton Press,  1989
>
>Nucleic acid hybridization:  A practical approach; 
>Hames BD and Higgins SL:  Oxford IRL Press,  1985
>
>PCR Protocols.  A Guide to Methods and Application;  Innis MA, Gelfand
>DH, Sinsky JJ and White TJ (editors):  Academic Press, Inc., 1990
>
>Molecular cloning:  A laboratory manual;  
>Maniatis TM, Fritsch EF and Sambrook J:  Cold Spring Harbor Laboratory,
>1982
>
>Fundamentals of Immunology;  
>Myrvik Q and Weiser R:  Lea & Febiger,1984
>
>Electron Microscopy in Molecular Biology;  
>Summerville J and Scheer U:  IRL Press, 1987
>Laboratory Manual of Histochemistry; Vacca LL:   Raven Press,  1985
>
>
>
>
>
>Manual of Immunoperoxidase Techniques;  
>Wordinger RJ, Miller GW, and Nicodemus DS:  ASCP Press, 1984
>
>"Immunohistochemistry" Polak, Julia M. & Van Noorden, Susan 
>  2nd ed. 1997. Bios Scientific Publishers, Oxford. 160 pages.
>
>Immunologic Staining, Electron Microscopy, and an Introduction to
>In-Situ Hybridization and Flow Cytomentry,  A Self-Assessment Exam;
>Elias JM, Council-Maguire N and Swisher B (editors)  
>Published by:National Society for Histotechnology  
>5900 Princess Garden Parkway  Suite 805  Lanham, MY 20706
>
>Ramos-Vara JA and Beissenherz ME, J Vet Diean Invest 12:307-311, 2000
>
>	>>>BIOS Scientific Publishers Limited 1999 aka Springer Verlag New
>York Inc  
>		>>>
>		>>>ISBN #0-387-91594-x
>		>>>
>	>>>Immunoenzyme Multiple staining Method,C.M. van der Loos 
>		>>>
>	>>>Amazon.com should also have it, or access it.
>		>>>
>>>>Try website , www.springer-ny.com <http://www.springer-ny.com>    
>
>NCCLS Quality Assurance for Immunocytochemistry: Proposed Guideline
>(NCCLS document MM4-P)
>
>HistoMark Double Staining Procedures.  #ML-168-02.  KPL 1-800-638-3167
>
>Handbook in Immunohistochemistry:  DAKO Corporation
>
>Nonradioactive in Situ Hybridization Application Manual, second edition,
>1996 Boehringer Mannheim Corporation 9115 Hague Road
>P.O. Box 50414 Indianapolis, IN 46250-0414 USA 1-800-428-5433
> 
>Journal of Histochemistry/Cytochemistry
>
>Journal of Histotechnology
>
>
>Cell Vision
>
>Analytical Morhphology, theory, applications and protocols
>Ed. Jiang Gu, 1997 Eaton Publishing Co. ISBN # 1-881299-03-1
>
>Immunogold Silver Stainng: principles, methods and applications
>Ed. M A Hayat, CRC Press, 1995 ISBN# 0-8493-2449-1
>
>OTHERS:
>
>Manual of Immunoperoxidase Techniques.  Wordinger RJ, et al
>ASCP Press ISBN# 0-89189-173-0
>
>Handbook, Immunochemical Staining Methods, DAKO Corp, Ed. Sally Naish, 1989
>Copyright of DAKO Corp, Carpentiera CA
>
>Applied Immunohistochemistry.
>ISSN 1062-3345
>Published quarterly by Lippincott Williams&Wilkins.
>US subscriptions Individual $125.00, Institution $156.00
>Editor-in-Chief: Clive Taylor
>Associate editors: Battifora, Cote, Gown, Rosai, Weiss.
>Official Publ of the Society for Applied Immunohistochemistry.
>
>
>
>
>
>Immunohistochemistry II.   A.C. Cuello  John Wiley & Sons, 1993
>
>Avidin-Biotin Chemistry: A Handbook.  Pierce Chemical Company, 1992
>
>Neil Hand's article, in Journal of Histotechnology
>on "Using Superheating using pressure cooking: its use and application
>in unmasking antigens embedded in  methylmethacrylate" 21:231-236, Sept 1998
>
>USEFUL INTERNET SITES FOR IMMUNOHISTOCHEMISTRY
>
>www.ihcrg.org <http://www.ihcrg.org>         NSH IHC Resource Group website
>
>http://www.medmatrix.org/index.stm/  Medical Matrix: A very useful
>guide to medical resources on the net
>
>http://www.ncbi.nlm.nih.gov/PubMed/  The National Library of
>Medicine's free search service
>
>http:igm.nlmgov/  Internet Grateful Med:  another National Library of
>Med. Free search service
>
>http://www.avicenna.com/  an excellent free MEDLINE/MEDLARS database
>search engine
>
>http://immuno.hypermart.net/  IMMUNOHISTOCHEMISTRY HOME PAGE:  useful
>IHC links
>
>http://abtx.wustl.edu/antibody.html#practive  IHC made simple, a nice
>general discussion of IHC
>
>See also:  Turbett GR and Sellner LN:  A comprehensive guide to
>pathology resources avalable on the internet.  Advances in Anatomic
>Pathology 3(5):313-335, 1996
>
>
>		-----Original Message-----
>		From:	Laura Collins [mailto:collinstralee@yahoo.com]
>		Sent:	Sunday, April 01, 2001 3:52 AM
>		To:	histonet@pathology.swmed.edu
>		Subject:	
>
>		Can anyone out there recommend recent books on
>		immunohistochemistry, both from a pathology point of
>		view and a laboratory technical aspect?  They would
>		need to include details on the uses of antibodies in
>		pathology reporting ie cytokeratins, lymphomas, tumour
>		markers, etc.  We are about to start this in our own
>		laboratory and need to access some recent
>		publications.
>
>		I would need to know the title, author(s), publisher,
>		publication year and if possible ISBN number.  
>
>		Any information would be appreciated.
>
>		Mark Collins
>		Senior Laboratory Technician
>		Tralee General Hospital
>
>
>		__________________________________________________
>		Do You Yahoo!?
>		Get email at your own domain with Yahoo! Mail. 
>		http://personal.mail.yahoo.com/?.refer=text
>
>
>----------------------------------------------------------------------
>
>Date: 2 Apr 2001 11:31:21 -0600
>From: Patsy.Ruegg@UCHSC.edu
>Subject: FW: Solochrome Azurine for AL
>
> 
>- -----Original Message-----
>From: Ruegg Patsy 
>Sent: Monday, April 02, 2001 10:48 AM
>To: 'Patty Lott'
>Subject: Solochrome Azurine for AL
>
>
>I purchased this stain from a German Company i believe, I think it was
>distributed by ROBOZ
>Pfaltz & Bauer, Inc.
>Solochrome Azurine, C.I. No. 43830
>Cat. No. S07772
> 
>good luck, it is not easy to find
> 
>Patsy Ruegg
>
>- -----Original Message-----
>From: Patty Lott [mailto:Patty.Lott@ORTHO.UAB.EDU]
>Sent: Monday, April 02, 2001 9:29 AM
>To: 'Patsy.Ruegg@UCHSC.edu'
>Subject: RE: stain for aluminum in plastic sections
>
>
>
>Thanks so much! 
>
>Say, where did you buy the solochrome azurine?  (I don't see it in Aldrich,
>which is my usual.) 
>
>- -----Original Message----- 
>From: Patsy.Ruegg@UCHSC.edu [ mailto:Patsy.Ruegg@UCHSC.edu
><mailto:Patsy.Ruegg@UCHSC.edu> ] 
>Sent: Friday, March 30, 2001 7:48 PM 
>To: Patty.Lott@ORTHO.UAB.EDU; Patsy.Ruegg@UCHSC.edu 
>Cc: histonet@pathology.swmed.edu 
>Subject: RE: stain for aluminum in plastic sections 
>
>
>Patty, 
>my AL procedure is very simple 
>make up a 0.5% solution of solochrome azurine in dih20 
>ph to 5 with weak acetic acid i believe (remind me to check this) 
>the solution will turn red (filter solution) 
>stain the gma sections in this solution for 1 to 12 hours at RT 
>pour off the stain and rinse very well in dih20 
>pour on a solution of 0.4M EDTA ph 7.4 for 30 min. 
>rinse well with dih20 
>pour on another change of 0.4M EDTA ph 7.4 for 30 min. 
>rinse well in dih20 and then let sit in dih20 for 5 min. until clear 
>rinse again 
>ary dry 
>coverslipp with permount 
>results: 
>AL will appear as blue deposit lines around bone 
>it is much like tetracycline in that it goes to where new bone is being 
>formed and deposits there, it is much more detramental than tet. though as 
>it prevents the formation of anymore bone after it is deposited, so it must 
>be chelated or the bones go to hell 
>
>Iron can do the same thing and deposit at the same mineralization front as 
>AL, so you must do an iron stain at the same time to confirm that the AL 
>positive reaction is really AL and not iron.  The iron stain does not stain 
>AL.  We published a paper on this a few years ago in JOH, I will find the 
>reference for you.  
>
>- -----Original Message----- 
>From: Patty Lott 
>To: 'Patsy.Ruegg@uchsc.edu' 
>Sent: 3/30/01 9:51 AM 
>Subject: stain for aluminum in plastic sections 
>
>Could I please get a copy of your procedure?  I'd love to try it!  (I've 
>already ordered the reagents for the other, but if this is much 
>better--which is what I am hearing--I need to know it!) 
>
>Thanks! 
>
>Patty Lott 
>Lab.Supervisor 
>Orthopaedic Research Laboratory 
>University of Alabama at Birmingham 
>
>
>
>----------------------------------------------------------------------
>
>Date: 2 Apr 2001 12:51:17 -0600
>From: "Su, Phy-Huynh" <psu@shctampa.usf.edu>
>Subject: RE: UNSUBCRIBE NOW!!!!! UNSUBSCRIBE NOW!!!!!! UNSUBSCRIBE NOW SEN	D
>No	More Messages
>
>
>
>> -----Original Message-----
>> From:	ACKAH, LILLIE M.(VSDP) [SMTP:ACKAH@mx.afrri.usuhs.mil]
>> Sent:	Friday, March 30, 2001 11:42 AM
>> To:	histonet@pathology.swmed.edu
>> Subject:	UNSUBCRIBE NOW!!!!! UNSUBSCRIBE NOW!!!!!! UNSUBSCRIBE NOW
>> SEND No	More Messages
>> 
>> To successfully unsubscribe, you have to type just 
>> Unsubscribe
>> in the subject line.  Don't type in anything else which will confuse the
>> system and you won't be removed from the list.
>> 
>> 
>
>
>----------------------------------------------------------------------
>
>Date: 2 Apr 2001 13:08:08 -0600
>From: Jianyun.Yin@aventis.com
>Subject: Histopath. scientist position
>
>Hi every one,
>
>The histopathology section in Respiratory and RA Disease Group in
>Aventis is Currently seeking a Contract Scientist for 6 months basis
>with the good possibility to transfer to regular employment. 
>
>The responsibility for this position is including:  performing routing
>histology procedures including paraffin and frozen tissue preparation,
>sectioning and staining, microscopy image collection and processing,
>development and application of immunohistochemistry and in situ
>hybridization protocols, interactive with other groups and project
>terms. 
>
>This individual must have an MS or BS and extensive research experiences
>in every aspect of histopathology work. Prior experienceaand applied
>knowledge of in situ Hybridization and immunohistochemistry. The
>candidate must have excellent
>communications and problem-solving skills.
>
>Please send your resume to MR. George Steidle in Aventis HR, Route
>202-206,
>Bridgewater, NJ 08807, georg.steidle@aventis.com
>
>or Call Jian Yin (908)231-4554
>
>
>
>
>
>
>
>
>
>
>
>
>
>
>
>  
> 
>
>
>
>
>
>
>
>----------------------------------------------------------------------
>
>Date: 2 Apr 2001 13:12:52 -0600
>From: Barbara.Davies@memhospcs.org
>Subject: Histotech opening
>
>Memorial Hospital in beautiful Colorado Springs, Colorado has a position
>open for a histotech.  Memorial processes 18,000 surgical pathology cases
>per year, working with state-of-the-art equipment and a great team of 6
>pathologists, 6 histotechs and 2 lab assistants.   Outstanding benefits and
>a competitive salary are offered.  HT or HTL(ASCP)  required.
>
>www.memorialhospital.com
>
> or contact:
>Memorial's jobline at 719-365-5113 or employment at 719-365-2335
>
>or email:
>Barbara Davies, Histology Manager
>barbara.davies@memhospcs.org
>
>
>
>
>----------------------------------------------------------------------
>
>Date: 2 Apr 2001 13:13:07 -0600
>From: conmac@cc.usu.edu
>Subject: Re: Is it formalin or saline
>
>That is a cool idea!  I will keep this in mind. We have a mystery
>solution that seems to appear from time to time in the tissue containers
>that are set in the necropsy/cutting room pass-thru.  It seems to be 70%
>ETOH judging from the way the tissues look when they're taken out for
>trimming in, but no one can figure out who puts it there or why.  We
>have such a small group of people working here, that it's usually fairly
>easy to track down who did what when, but not this.  anyway, this little
>trick.. and the one John Kiernan mentions using Schiff reagent, will
>come in handy when this happens again.
>
>Connie M.
>
>
>David Grehan wrote:
>> 
>> Have you ever been asked if a specimen has been placed in formalin or
>> saline but you are reluctant to have a sniff and blast your mucous
>> membranes?.
>> Here is a simple test ;
>> Add  few drops of the fluid to 1 ml of your reticulin silver solution.
>> If it turns white the fluid is saline.
>> If it turns black you have got formalin .
>> 
>> Easy!
>> 
>> David Grehan
>> .
>
>- -- 
>D#236##017#a#193##177##026#a
>
>
>----------------------------------------------------------------------
>
>Date: 2 Apr 2001 14:47:59 -0600
>From: HistoHacker <histohacker@yahoo.com>
>Subject: Apoptosis staining
>
>Hello Histoland,
>One of my docs needs a protocol for a Hoerst (sp?)
>stain (for apoptosis).  Can anyone help?  Thanks in
>advance.
>Have a great day,
>Rebecca
>
>__________________________________________________
>Do You Yahoo!?
>Get email at your own domain with Yahoo! Mail. 
>http://personal.mail.yahoo.com/?.refer=text
>
>
>----------------------------------------------------------------------
>
>Date: 2 Apr 2001 14:48:21 -0600
>From: "Vinnie Della Speranza" <dellav@musc.edu>
>Subject: TEST MESSAGE, again  please ignore
>
>
>
>----------------------------------------------------------------------
>
>Date: 2 Apr 2001 15:20:37 -0600
>From: Andrea Grantham <algranth@u.arizona.edu>
>Subject: web site
>
>Histonetters,
>My boss asked me to post the following message to Histonet:
>
>
>Dear Colleagues,
>I am pleased to announce the completely revised and updated edition of
>"Microscopy & Imaging Resources on the WWW" is now available at a new
>URL. The goal of this site has always been to provide resources for
>University students, staff and faculty who want to learn more about
>microscopy and imaging. The site includes K-12 educational links,
>information on Light Microscopy, Histology, Confocal Microscopy,
>Fluorescence Techniques, Electron Microscopy, and Digital Imaging.
>http://swehsc.pharmacy.arizona.edu/exppath/
>"Microscopy & Imaging Resources on the WWW" is an outreach of the Southwest
>Environmental Health Sciences Center, which is funded by the National
>Institute of Environmental Health Sciences (NIEHS is one of the National
>Institutes of Health, part of the U.S. Department of Health and Human
>Services).
>Yours,
>Doug Cromey
>Manager, Experimental Pathology Core, SWEHSC
>
>
>.....................................................................
>: Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy :
>: Research Specialist, Principal University of Arizona :
>: (office: AHSC 4212A) P.O. Box 245044 :
>: (voice: 520-626-2824) Tucson, AZ 85724-5044 USA :
>: (FAX: 520-626-2097) (NEW email: Cromey@Arizona.edu) :
>:...................................................................:
>http://swehsc.pharmacy.arizona.edu/exppath/
>Home of: "Microscopy and Imaging Resources on the WWW
>
>
>
>
>.....................................................................
>: Andrea Grantham, HT(ASCP)     Dept. of Cell Biology & Anatomy     :
>: Sr. Research Specialist       University of Arizona               :
>: (office:  AHSC 4212)          P.O. Box 245044                     :
>: (voice:  520-626-4415)        Tucson, AZ  85724-5044    USA       :
>: (FAX:  520-626-2097)          (email:  algranth@u.arizona.edu)       :
>:...................................................................:
>           http://www.cba.arizona.edu/histology-lab.html
>
>
>
>----------------------------------------------------------------------
>
>Date: 2 Apr 2001 15:59:41 -0600
>From: "Dr. Allen A. Smith" <asmith@mail.barry.edu>
>Subject: Re: Ring-around-the-collar.
>
>Paddington is a bear in a series of children's stories.  Since he is
>cleverer (but no wiser) than Winnie-the-Pooh, he wears wellies to keep his
>hindpaws dry.
>Wellies are short for "Wellingtons", mid-calf length boots.  Children's
>wellies are usually made of black rubber and leave a ring-around-the-calf if
>worn with shorts.
>(When I lived in Philadelphia, I wore leather wellies in in winter.)
>
>Allen A. Smith, Ph.D.
>Barry University  School of Graduate Medical Sciences
>    Podiatric Medicine and Surgery
>Miami Shores, Florida
>
>- ----- Original Message -----
>From: Connie McManus <conmac@cc.usu.edu>
>To: Dr. Ian Montgomery. <ian.montgomery@bio.gla.ac.uk>
>Cc: <histonet@pathology.swmed.edu>
>Sent: Friday, March 30, 2001 8:06 PM
>Subject: Re: Ring-around-the-collar.
>
>
>Dr. Ian,
>
>Huh???  Please explain what Paddingon and Wellies are.
>
>thnx
>Connie M
>
>"Dr. Ian Montgomery." wrote:
>>
>> John,
>>         What about the other ring, common in children from the UK, the
>> ring-around-the-calf. Mostly found in Primary School children during
>> the long dark wet winter months when Wellies are a common footwear. As
>> you'll probably remember, the ring is an almost constant feature among
>> children who develop a love for Paddington and have to be surgically
>> removed from the beloved Wellies.
>>         I wonder if Compo went to his grave with the ring, he almost
>> certainly had one around-the-collar.
>>  Ian.
>>
>> Dr. Ian Montgomery,
>> West Medical Building,
>> University of Glasgow,
>> Glasgow,
>> G12 8QQ.
>> Tel: 0141 339 8855.  Extn:6602.
>> Fax: 0141 330 2923
>> e-mail: ian.montgomery@bio.gla.ac.uk
>
>- --
>-oe#017##224##161##177##026##225#
>
>
>
>
>
>
>----------------------------------------------------------------------
>
>Date: 2 Apr 2001 16:51:36 -0600
>From: Patsy.Ruegg@UCHSC.edu
>Subject: RE: Apoptosis staining
>
>Do you mean the DNA Hoechst Assay?
>I don't think this will really help you determine apoptotic cells but if you
>want the procedure I could fax it to you if I had your fax number.  Sorry
>don't have it on my computer for emailing.  Hey I just remembered I have a
>new machine at home that will scan it.
>Patsy Ruegg
>
>		-----Original Message-----
>		From:	HistoHacker [mailto:histohacker@yahoo.com]
>		Sent:	Monday, April 02, 2001 1:14 PM
>		To:	histonet@pathology.swmed.edu
>		Subject:	Apoptosis staining
>
>		Hello Histoland,
>		One of my docs needs a protocol for a Hoerst (sp?)
>		stain (for apoptosis).  Can anyone help?  Thanks in
>		advance.
>		Have a great day,
>		Rebecca
>
>		__________________________________________________
>		Do You Yahoo!?
>		Get email at your own domain with Yahoo! Mail. 
>		http://personal.mail.yahoo.com/?.refer=text
>
>
>----------------------------------------------------------------------
>
>Date: 2 Apr 2001 17:09:02 -0600
>From: "Vinnie Della Speranza" <dellav@musc.edu>
>Subject: Re: Muscle Biopsy Clamps
>
>Greg,
> 
>thank you for sharing your muscle biopsy protocol with us. I am in complete
>agreement with your remarks regarding the value of requesting sufficient
>samples at biopsy.
> 
>you may be interested to know that I have achieved acceptable EM results by
>using 4% paraformaldehyde fixative which has allowed us to get by with one
>clamped specimen, in addition to the unfixed sample for enzyme
>histochemistry.
>This fixative eliminates the methanol contained in commercial
>formalin/formaldehyde solutions that can compromise EM. You may experience
>less resistance from clinicians if you can get by with only two samples.
>Vinnie
> 
>
>Vinnie Della Speranza
>Manager for Anatomic Pathology Services
>Medical University of South Carolina
>165 Ashley Avenue
>Suite 309
>Charleston, SC  29425
>ph:  (843) 792-6353
>fax: (843) 792-8974
>
>
>>>> <GregorLuck@aol.com> 03/31/01 08:57PM >>>
>Peggy et. al.,
>Our protocol (from Dr. Sumi @ U. of W. neuropath) requests the following for 
>optimal
>neuropathathology workup on a muscle biopsy:
>       1.  One biopsy "unclamped" for muscle enzyme histochemistry (Note, for
>the           reasons of clamp artifact and the desired uncrushed relaxation 
>of  the fibers, as you noted.)  This portion is snap frozen in a relaxed 
>state @ -160 degrees C. in  Isopentane cooled by liquid nitrogen.
>       2.  A 2nd biopsy in a clamp and allowed to fix in this state in 10% 
>NBF for 24 hours before resection of the "uncrushed" portion of the biopsy 
>from between the tines of the clamp(for routine L.M. if needed).
>       3.  A 3rd biopsy (as in #2 above) but in Trump's fixative for E.M. if 
>needed.
>Our feeling is that if the patient is going to undergo an open biopsy 
>procedure we should harvest all materials necessary for a complete and 
>definitve neuropathology assessment.  The only time we compromise on these 
>specimen requirements are when the patient is a pediatric or a Shriner's 
>patient when there is concern for the amount of the muscle compromise to the 
>young patients.  I understand that often not all three of these specimens are
>necessary for diagnosis, but after getting the patient all the way into an 
>invasive surgical procedure, which is for the sole purpose of obtaining a 
>tissue diagnosis,  I would advocate a multiple bx/fixative approach.
>Greg Luck
>Anat.Path. Sup.
>Deaconess Med Center
>Spokane, WA
>
>
>
>----------------------------------------------------------------------
>
>Date: 2 Apr 2001 17:09:23 -0600
>From: lbrennan@coh.org
>Subject: Thermanox Coverslips
>
>So sorry, I was responding to the graduate student doing ISH on cell
>cultures.  Not all plastic coverslips dissolve in acetone or xylene!
>     Lynn Brennan
>
>
>
>----------------------------------------------------------------------
>
>Date: 2 Apr 2001 20:34:44 -0600
>From: Patsy.Ruegg@UCHSC.edu
>Subject: hoechst dna stain
>
> 
>
>- -----Original Message-----
>From: rueggp
>To: patsy.ruegg@uchsc.edu
>Sent: 4/2/01 7:44 PM
>Subject: dna stain
>
>Reference: Cesar La Barca and Kenneth Paigen
>A Simple~ Rapid~ and Sensitive DNA Assay Procedure. Analytical Biochern
>102, 344-352.
>I. Stock Solutions
>A. Hoechst Dye Stock Concentrate
>0.5 mg/mi Hoechst dye in water.
>Store protected from light, at 4#161#C, for up to 6 months. Working
>Solution: 4 ul in 10 mi 4.6 M NaCI-PO4 buffer. Make up fresh before each
>use.
>B. DNA STANDARD STOCK
>5 mg calf thymus DNA in 10 mi 5 mM NaOH
>Allow to stir overnight at 4#161#C, pipet to further dissolve. Aliquot into
>10 mi sterile tubes (200 ul) and store at -20#161#C.
>C. 4.6 M NaCI-PO4 buffer
>1.14 g/L NaH2PO4 (anhydrous)
>5.753 g/L Na2HPO4 (anhydrous) 268.6 g/L NaCI
>Adjust pH to 7.4
>D. 1M NaOH
>E. 1M HCI+200mM PO4
>500 mi X (50 mM NaH2PO4)
>(150 mM Na2HPO4) Adjust pH to 7.4
>Discard 41.67 mi of buffer, adjust volume to 500 mls with 12 M HCI.
>VERY IMPORTANT
>It is now necessary to ensure that the NaOH and
>1M HCI+200mM PO4 are of equal molarities. To a 50 ml tube, add 5 mis of
>each, and check the pH. If other than pH 7.4 adjust concentrations until
>result of the addition is 7.4.
>F. 0.25 M NaCI
>
> ." ".;."'..""c."""'",..,,
>II. CELL PREPARATION
>A. Aliquot 100 ul sample ~to 1.5 mI tubes. Add 200 ul
>1 M NaOH. Vortex well and freeze at -20#161#C. Incubate 1 hr at room
>temperature, or overnight at 4#161#C.
>B. Add 200 ullM HCI+200mM PO4. Sample is now ready.
>III. PREPARATION OF DNA STANDARD CURVE A. DNA SOLUTION A
>Remove 1 tube of DNA stock solution from -20#161#C freezer. Add 1.9 mIIM
>HCI+200mM PO4 and 1.9 mI NaOH. B. DNA SOLUTION B
>Tol.6 mI 0.25 M NaCI, add 400 ul DNA SOLUTION A.
>IV .DNA ST ANDARD CURVE
>Add directly to microflour 96 well plate the following amounts:
>DNA final concentration 0.25 M NaCI DNA A ~NA B
>3.2 ug/mI 93 ul 32 ul
>1.6 ug/mI 109 ul 16 ul 0.8 ug/mI 117 ul 8 ul
>0.4 ug/mI 105 ul 20 ul 0.2 ug/mI 115 ul 10 ul 0.1 ug/ml 120 ul 5 ul 0.0
>ug/mI 125 ul
>You must load plate in dupicate only, starting from highest
>concentration and going to lowest, in 125 ul volumes.
>A. Add 125 ul of sample into remaining wells, in duplicate.
>B. Add 125 ul Hoechst dye (of working concentration) to each well.
>Incubate in light protected environment, for at least 20, but less than
>60 minutes
>C. Read on Dynatech Microflour plate reader for relative
>flourescence units. Estimate the DNA content of samples from standard
>curve.
>
>
>
>
>----------------------------------------------------------------------
>
>Date: 2 Apr 2001 21:48:04 -0600
>From: Victoria Baker <vbaker60@yahoo.com>
>Subject: NFkBeta - p50, p52 & p65 in paraffin (FFPE) sections
>
>
>  
>
>
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