Hi Histonetter
		Is any body working on Tendons & Ligament ?? I am interested
in 1) What is the best way to fix & process in order to perform Immuno or
Routine . Please reply to me at Shakun.Aswani@Roche.com Thank you very much
Shakun

> -----Original Message-----
> From:	HistoNet Server [SMTP:HistoNet@Pathology.swmed.edu]
> Sent:	Tuesday, December 01, 1998 10:03 PM
> To:	HistoNet Server
> Subject:	Daily Digest
>
>
> ----------------------------------------------------------------------
>
> Date: 1 Dec 1998 00:08:24 -0600
> From: "Paul Marr" <pmarr@ventanamed.com>
> Subject: ATTN: So. California Society for Histotechnology
>
> Histonetters:
>
> Ventana Medical Systems will be hosting an Open House on 02 DEC at the San
> Diego Doubletree Hotel in Mission Valley from 11:00 A.M. until 7:00 P.M.
> for
> the launch of our automated Special Stains System in conjunction with the
> CA
> Society for Histotechnology, San Diego Chapter.  Dr. Rachael Morecki will
> give a lecture on Colon cancer at the Meeting @ 4:00 P.M.  All
> Pathologists,
> Histology Technologists, Lab Managers and Administrators who can attend
> are
> welcome.  There will be a "Hands-On" Demonstration of our Special Stains
> System as well as reimbursement information for managers and
> administrators.
> The San Diego Doubletree Hotel is located at 7450 Hazard Center Drive, San
> Diego and the phone number is: (619) 297-5499.  Thank you.
>
> Paul Marr
> Account Executive
>
>
>
> ----------------------------------------------------------------------
>
> Date: 1 Dec 1998 00:08:47 -0600
> From: Michelle Down <michellD@qimr.edu.au>
> Subject: RNA
>
> Hi again,
> Does anyone have a method for extracting total RNA from formalin fixed
> cells (ie peripheral blood lymphocytes). Or another method for fixing
> cells
> that preserves RNA for extraction?
> Thanks
> Michelle
>
>
>
> ----------------------------------------------------------------------
>
> Date: 1 Dec 1998 11:57:31 -0600
> From: Cynthia Favara <cfavara@atlas.niaid.nih.gov>
> Subject: enclosure check
>
> Just checking to see if we have fixed our enclosure problem at this end.
> Please let me know. Thanks
>
> Cynthia Favara
> 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: 1 Dec 1998 11:57:55 -0600
> From: andrea_kelly@CCGATEWAY.AMC.EDU (by way of Histonet)
> Subject: Re: What pressure is needed on a vacuum oven?
>
>       I recall that it should pull 20-25 mgHg.
>
>       Andrea Kelly
>
>
> ______________________________ Reply Separator
> _________________________________
> Subject: What pressure is needed on a vacuum oven?
> Author:  Karen S Pawlowski <kna101@utdallas.edu> (by way of Histonet) at
> Internet-Mail
> Date:    12/1/98 7:25 AM
>
>
> Hi all,
>
> We recently inherited a vacuum oven for hand processing paraffin blocks.
> Actually, I don't know what the oven was originally used for, but we want
> to use it to process paraffin blocks.
> It can be set at any pressure we want, but I don't know the pressure that
> is needed for parafin processing and I can't find it listed in any of my
> histology texts.  Anyone out there know what it is off hand?
>
> Thanks in advance,
>
> Karen Pawlowski
> Dept. of Otolaryngology
> UT Southwestern Med. Ctr.
> Dallas, TX
>
>
>
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 1 Dec 1998 11:58:17 -0600
> From: "Hinton, Sandy" <sahinton@utmb.edu> (by way of Histonet)
> Subject: IHC Disclaimer
>
> This may be a subject that has been discussed previously but if so I was
> off
> the network at that time.
> I have a copy of the Federal Register relating to the ruling on
> Immunohistochemistry reagents.
> My question is what disclaimer is CAP requiring us to add to final
> Surgical
> Pathology reports, to address Immunohistochemistry procedures?
> Please feel free to email or FAX (409) 772-4676, your response.
> Thanks
> University of Texas Medical Branch at Galveston
> Sandy Hinton
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 1 Dec 1998 11:58:52 -0600
> From: "D. Hammer" <hammerd@u.washington.edu> (by way of Histonet)
> Subject: Re: Alcohol volumes
>
> Hey, who is this "Masked Man"??  He sure has a bead on flammable storage.
>
> Thanks for the great layout of info.
>
> Don
>
> ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
> Don Hammer, Administrative Director            UNIVERSITY OF WASHINGTON
> Hospital Pathology, Box 356100                     MEDICAL CENTER
> 1995 NE Pacific St.
> Seattle Washington, 98195                  ~Where Knowledge Comes To Life~
> (206) 548-6401 Fax: (206) 548-4928
> ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
>
>
> On Mon, 30 Nov 1998, Mathew Osborn wrote:
>
> > Histonetters,
> >
> > Recently a question was posed regarding the allowable volumes of alcohol
> > in a lab.  I hope what follows sheds some light on this issue.
> >
> > Ethanol is classified as a Class IB combustible liquid. Class IB liquids
> > have a maximum allowable container capacity of 20 liters in "metal (non
> > DOT
> > approved) or approved plastic containers, safety cans or DOT spec metal
> > drums."
> >
> > The amount of combustible liquids allowable is determined by the size of
> > the "laboratory unit". For hospitals with sprinkler systems, per 100 sq.
> > feet of lab space, Class I, II and IIIA (including IB) flammable liquids
> > are limited to 4 L (1.1 gallons) not in approved storage containers or
> > safety cans and 7.5 L (2 gal) total including quantities in storage
> > cabinets and safety cans. That is, only half of the combustibles in a
> > lab
> > can be outside of approved storage containers. Note that there is no
> > rule
> > governing total volume except as it pertains to lab area and to
> > container
> > size.
> >
> > The key definition here is "laboratory unit". Some EH&S officers define
> > laboratory unit roughly as the space enclosed by fire doors. Thus, for
> > all
> > but the smallest hospitals and labs, there should be few restrictions
> > because of space-related alcohol volume limitations. As an example, one
> > Sakura VIP has 21 L of ethanol and 9 L of Xylene (Class IIIA). Assuming
> > that a tissue processor is not an approved storage container, this means
> > that the "lab unit" would need to be a minimum of 750 sq. ft. (30x25
> > ft.)
> > for each tissue processor to be "legal" ((30 L o 4 L) X 100 sq. ft. ).
> > For
> > a tissue processor that is considered to be an approved storage
> > container,
> > then the space requirements are halved.
> >
> > I hope this helps to answer the question.
> >
> > If you have concerns or questions about cost effectively recycling these
> > solvents, please contact me.
> >
> > Sincerely,
> >
> > Matt Osborn
> > Product Manager
> > Naiad Technologies, Inc.
> > osborn@naiadtech.com
> > 503-274-4407
> >
> >
> >
> >
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 1 Dec 1998 11:59:15 -0600
> From: Gayle Callis <uvsgc@msu.oscs.montana.edu> (by way of Histonet)
> Subject: pressure/hand processing/vacuum oven
>
> We draw a vacuum to 15 - 20 psi, make sure the thermometer is visible
> with temp set at 60C or so. You can mimic a vacuum pressure automated
> processor by pulling the vacuum, releasing it after a specified time
> then draw the vacuum again.  Voila!  Alternating vacuum pressure,
> with good infiltration!  This can be improve dense tissue infiltration.
>
> Gayle Callis
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 1 Dec 1998 11:59:54 -0600
> From: "Sebree Linda A." <la.sebree@hosp.wisc.edu> (by way of Histonet)
> Subject: RE: Plap
>
> Dear Alex,
>
> We ran into the same problem early on; the placenta control was intensely
> positive but known germinomas were dead negative.  We no longer use
> placenta
> as a control but rather known positive tumors.  The dilution used for
> placenta was not nearly strong enough to stain known positive tumors so
> now
> we optimize our PLAP protocol using germinomas.  I'm certain this will
> solve
> your problem.
>
> Linda
>
> P.S.  Just looked up our optimization notes: we used Dako's polyclonal at
> 1:3200 for placenta but reoptimized down to 1:100 for
> germinomas/seminomas!
>
> Linda Sebree, HT
> University of Wisconsin Hospital & Clinics
> Department of Laboratory Medicine
> IHC/ISH Laboratory
> A4/204-2472
> 600 Highland Ave.
> Madison, WI  53792-2472
>
> Phone:  (608)265-6596
> FAX:  (608)263-1568
>
> > -----Original Message-----
> > From:	Alex Brown [SMTP:AlexB@nayrshire.scot.nhs.uk]
> > Sent:	Monday, November 30, 1998 11:06 AM
> > To:	'Histonet'
> > Subject:	Plap
> >
> > Hi All,
> >           We've recently been having some trouble with our Placental
> > Alkaline Phosphatase (Plap).  i.e. variable results. Some tumours we
> > expected to be positive, have been negative although the control (
> > normal placenta) has been strongly positive. In one case recently, a
> > tumour in testes (well fixed), allthough the tumour cells were
> > unexpectedly negative, there was positive staining in smooth muscle
> > fibres.
> > 	We use Dako's Primary (M7191, Clone 8A9) at 1:50 for 30mins ,
> > microwave/pressure cook, and use Biogenix's HRP Kit and DAB. Staining is
> > carried out on the Optimax II.
> > 	Anyone else had any similar problems with this ??????
> > 		Regards
> > 			Alex. Brown
> > 			Crosshouse Hospital
> > 			Kilmarnock, Scotland.
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 1 Dec 1998 12:00:24 -0600
> From: "Weems, Joyce" <JWEEMS@sjha.org> (by way of Histonet)
> Subject: FW: bcl-2
>
> I am resending this from last week because I've not had any response!
> J:>)
> >----------
> >From: 	Weems, Joyce
> >Sent: 	Friday, November 27, 1998 9:34 AM
> >To: 	'Histonet'
> >Subject: 	bcl-2
> >
> >Hey you experts!
> >
> >What have you found to be the best method of determining overexpression
> of
> >bcl-2 and bcl-1? Specimen type, etc. PCR, Immunoperoxidase, etc?
> >
> >Thanks for all your help!
> >Joyce
> >
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 1 Dec 1998 12:00:53 -0600
> From: jo-ann@lan1.molonc.mcgill.ca (Jo-Ann Bader) (by way of Histonet)
> Subject: Tissue Culture help
>
> I realise that this may not be the right place for tissue culture but if
> anyone out there knows of a web page or sight to post problems on tissue
> culture.  We have a PI who's projects have come to an almost complete halt
> because of dying cells.  Sorry I do not know much more about the problem
> but if I could supply her with an address of some sort that they could go
> to I'm sure it would be greatly appreciated.
>
> Thanks in advance
> Jo-Ann
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 1 Dec 1998 12:01:17 -0600
> From: Neuropathology Laboratory <neuropath.frenchay@dial.pipex.com> (by
> way of
> Histonet)
> Subject: Re:- PLAP problem
>
> Hi Alex,
> About a year ago we had similar problems with PLAP ( placental alkaline
> phosphatase). We tried the DAKO antibody that you are having problems
> with and found it quite reliable when used at a 1:10 dilution for 20mins
> after microwave antigen retrieval.
> We also tried the Biomen antibody ( monoclonal clone PL8-F6 )which was
> great. It does not require antigen retrieval and gave good results at the
> following dilutions:-  1:100  20mins incubation
>                        1:2000 overnight incubation
> We now use this antibody and (fingers crossed) don't appear to be having
> any problems.
> Our detection system is Vector Elite and DAB and our staining is carried
> out on a Leica Ig stainer.
> Please note that our immuno is carried out on Neuropathology specimens
> that have generally had less than 24hours fixation.
> Hope this is of help.
> Now can you help us? We are currently looking to replace our immuno
> stainer and would like your opinion on the Optimax II.
> Thanks in advance
> Sharron Williams
> Neuropathology
> Frenchay Hosp
> Bristol
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 1 Dec 1998 12:02:00 -0600
> From: "Instrumedics, Inc." <info@instrumedics.com> (by way of Histonet)
> Subject: cutting frozen chicken tendons
>
> Hi Kathy,
> Saw your message on the histonet and wanted to suggest that you might be
> helped with your chicken tendons with the CryoJane Tape-Transfer process.
> It
> is possible with CryoJane to section bone, tendons, fat and any other
> tissue
> without compression. The end result is a flat ,thin(as thin as 2 microns)
> section bonded to the slide. The section is supported and captured on a
> tape
> window as it is being cut. It is then transferred to an adhesive-coated
> slide which is then polymerized. This converts the coating to a hard
> plastic
> with the section bonded to it. The section will not dislodge no matter
> what
> rigorous protocol you use!
>
> This system can be installed in virtually all cryostats!
>
> If you are interested in the complete story please contact us. Also do
> visit
> our web site
>                                          www.instrumedics.com
>
> Bernice Schiller
> schiller@instrumedics.com
> 800-237-2772
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 1 Dec 1998 12:02:24 -0600
> From: "Sarah Christo" <schristo@CVM.TAMU.EDU> (by way of Histonet)
> Subject: What pressure is needed on a vacuum oven? -Reply
>
> Dear Karen,
>    Don't go over 15 pounds of vacuum
> (or inches of mercury).
> - -Sarah Christo, Texas A&M
>
>
> Hi all,
>
> We recently inherited a vacuum oven for hand processing paraffin blocks.
> Actually, I don't know what the oven was originally used for, but we want
> to use it to process paraffin blocks.
> It can be set at any pressure we want, but I don't know the pressure that
> is needed for parafin processing and I can't find it listed in any of my
> histology texts.  Anyone out there know what it is off hand?
>
> Thanks in advance,
>
> Karen Pawlowski
> Dept. of Otolaryngology
> UT Southwestern Med. Ctr.
> Dallas, TX
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 1 Dec 1998 12:03:06 -0600
> From: "Georger, Mary" <mary.georger@arcus.us.astra.com>
> Subject: RE: ChAT and Enkaphalin antibodies
>
> Joyce,
> I have used the Chemicon antibody on Formalin fixed rat tissue. If you are
> interested in the specifics let me know. It did require HIER.
> Mary Georger
>
>
> > ----------
> > From: 	Joyce Kotzuk[SMTP:JKotzuk@salud.unm.edu]
> > Sent: 	Monday, November 30, 1998 11:50 AM
> > To: 	HistoNet@Pathology.swmed.edu
> > Subject: 	ChAT and Enkaphalin antibodies
> >
> > Does anyone know of antibodies for ChAT (or acetylcholine neurons) and
> > met-enkaphalin that work well in paraffin embedded human autopsy tissue?
> I
> > have the Chemicon antibodies for these substances, but they only seem to
> > work for me using paraformaldehyde fixed, frozen sections. If anyone has
> > any suggestions for making these antibodies work in paraffin material,
> > that would be very appreciated as well. Thank-you.            Joyce
> Kotzuk
> >                                                                   Univ.
> of
> > New Mexico, Pathology
> >
> >
>
>
> ----------------------------------------------------------------------
>
> Date: 1 Dec 1998 18:55:00 -0600
> From: "Mick Rentsch" <ausbio@nex.com.au> (by way of Histonet)
> Subject: Re:- Alcohol limits and storage
>
> Dear colleagues,
> you will find there  is a plethora of regulation governing amounts,
> storage
> and conditions of storage: these will range from State to Federal and from
> authorities such as the Health Dept., Building Authority, OH&S, Transport
> Authorities etc. The bulk of these will be covered by some form of
> Legislation eg. Dangerous Goods Act., Poisons Act., Uniform Building
> Code.,
> Standards Assoc., Fire Act., and the list goes on.
> At any given time in your Institution any or all of the Legislations may
> apply as may do the Authorities; so, gather them all, select the most
> stringent and apply it.
> It is clear from some experienced persons that compliance is difficult if
> not damn impossible in some instances,it is therefore critical that proper
> representation be made to the relevant authorities etc. to implement
> change
> that allows for good work practice and sensible regulation.
> In my old lab, which was in an old bank building, we stored our flammables
> in the Vault, which was contructed out of three foot concrete/steel and we
> modified by incorporating bunding in the floor and putting an explosion
> panel in the roof vented to the outside; this certainly met our local
> regs.
> as having greater than a 2hr fire rating and stored less than 1000litres.
> The internal light was a flash-proof/ignition proof type. The new lab.
> since
> constructed has an external flammable store 5 meters from the Histo lab.
> and
> the entire Lab flammables are kept there, including Haematology, Micro and
> Biochem. Staff still use an internal Flamm. cupboard DGA Appd, but this
> only
> has a max. capacity of 100 litres, and in which we store only 60-70 Litres
> (enough for changing the processorsx2 ,stain machines and a flamm.waste
> drum)
> I would point out that I have seen several instances where in industry,
> Steel shipping containers have been converted into Flammable stores, and
> we
> had contemplated doing this but the Boss would have lost his car parking
> space (We didn't mind). Again these need to sited at least 5meters from a
> building and three meters from a clear title boundary.
> Common sense and innovation may allow you to stay within regs. but be
> prepared for the 5 meter walk and don't forget your raincoat.
> Regards Mike Rentsch (Downunder)
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 1 Dec 1998 18:55:24 -0600
> From: "Peter A. Takes" <ptakes@stereotaxis.wustl.edu> (by way of Histonet)
> Subject: Re: IHC Disclaimer
>
> Sandy:
>
> The disclaimer in the Federal Register notice will likely be required by
> CAP for
> tests where an ASR is used, in accordance with FDA requirements.  Please
> note
> that FDA now requires labs use that disclaimer, and labs employing ASRs
> now
> fall
> within FDA's jurisdiction for compliance with the report label
> regulations.
> See
> Clinical Laboratory News 24(4):10, April, 1998.
>
> Peter
>
> - --
> Peter A. Takes, Ph.D., RAC
> Director, Clinical & Regulatory Affairs
> STEREOTAXIS, Inc.
> Ph. 1-314-615-6964; Pager: 841-9351
> ptakes@stereotaxis.wustl.edu
>
>
> Hinton, Sandy (by way of Histonet) wrote:
>
> > This may be a subject that has been discussed previously but if so I was
> off
> > the network at that time.
> > I have a copy of the Federal Register relating to the ruling on
> > Immunohistochemistry reagents.
> > My question is what disclaimer is CAP requiring us to add to final
> Surgical
> > Pathology reports, to address Immunohistochemistry procedures?
> > Please feel free to email or FAX (409) 772-4676, your response.
> > Thanks
> > University of Texas Medical Branch at Galveston
> > Sandy Hinton
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 1 Dec 1998 18:55:49 -0600
> From: "Steven E. Slap" <ebs@ebsciences.com> (by way of Histonet)
> Subject: Re: Vibratome
>
> Dear Shirley and HistoNetters,
>
> I've been out of the country and I'm still sifting through my e-mails, so
> please forgive the delay in responding to this post.
>
> Shirley Chu asked about "Vibratomes."  "Vibratome" is a registered
> trademark of Technical Products International (TPI).  TPI have 5
> authorized distributors in the United States (we are one of them).
>
> Several other companies make vibrating blade microtomes, including Dosaka
> EM in Japan (Microslicers), Leica, EMS and ourselves (the "MicroCut").  I
> would encourage Shirley to compare the specifications of all the
> different models.
>
> Best regards,
> Steven E. Slap, Vice-President
>
>
>
> ********************************
> Energy Beam Sciences, Inc.
> The Laboratory Microwave Company
> http://www.ebsciences.com
> ********************************
>
>
>
>
> Here are the messages received yesterday!