[Histonet] RE:Gram stain (Mesru T) Histonet Digest, Vol 119, Issue 4

Hardy, Cate chardy <@t> csu.edu.au
Thu Oct 3 17:43:22 CDT 2013


We perform Gram Twort for our gram stains. It is essentially the same as the gram stain used in microbiology except that we counter stain with a neutral red fast green which is very effective.

Cheers Cate


Cate Hardy

Senior Technical Officer
Veterinary Diagnostic Laboratory
School of Animal and Veterinary Sciences
Locked bag 588
Charles Sturt University
Wagga Wagga
NSW 2678
T: 02 69334000
Email: chardy <@t> csu.edu.au
Email: vdl <@t> csu.edu.au




-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of histonet-request <@t> lists.utsouthwestern.edu
Sent: Friday, 4 October 2013 12:07 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 119, Issue 4

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Today's Topics:

   1. Gram stain (Mesru T)
   2. Benchmark Ultra troubles (Gudrun Lang)
   3. Re: RE: MOHS IPs (Kim Donadio)
   4. Re: IHC turnaround time (Kim Donadio)
   5. Re: Gram stain (Jennifer MacDonald)
   6. Hi (Helene Degan)
   7. RE: Validating new Benchmark instrument (Eytalis, Robert A)
   8. need lever clamp for blade of leica microtome (Patricia F Lott)
   9. Re: Shipping to Countries Outside of the US (Jan Shivers)
  10. RE: Histotechnologist, (ASCP, QIHC certified) and  Pathology
      assistant position in Seattle area (Yan Liang)
  11. Vacuum and pressure in tissue processing (Teri Johnson)
  12. Trivew (Gerald Davydov)
  13. Inconsistent Sections with Cryostat
      (Perow, Elliott S (PEROWES10))
  14. HT HistoDeck question... (Stephenson, Sheryl)
  15. RE: Inconsistent Sections with Cryostat  (Manfre, Philip)
  16. Re: HT HistoDeck question... (Lee & Peggy Wenk)
  17. Re: RE: Inconsistent Sections with Cryostat  (Lee & Peggy Wenk)
  18. RE: HT HistoDeck question... (Watson, Linda)
  19. RE: HT HistoDeck question... (Bernice Frederick)
  20. The NSH was AMAZING!!! Congratulations to the 2013
      Leadership,       Education and Advocacy Award Winners!! From Pam
      Barker and RELIA  Solutions!! (Pam Barker)


----------------------------------------------------------------------

Message: 1
Date: Wed, 2 Oct 2013 13:11:22 -0400
From: Mesru T <turkekul <@t> gmail.com>
Subject: [Histonet] Gram stain
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
        <CACf3QnTkAREiduT-jCBZ6CGbd6Bn0qudF4cM6FE1F9TVgYE5=A <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

Hi All,



Does anybody have a protocol for Gram stain on FFPE mouse intestine sections. We are looking to distinguish between Klebsiella and Vancomycin Resistant Enterococcus (VRE). I would like to avoid using Picric acid as some protocols suggest.

Thanks in advance.


------------------------------

Message: 2
Date: Wed, 2 Oct 2013 19:38:56 +0200
From: "Gudrun Lang" <gu.lang <@t> gmx.at>
Subject: [Histonet] Benchmark Ultra troubles
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <001a01cebf96$45a20a10$d0e61e30$@gmx.at>
Content-Type: text/plain;       charset="us-ascii"

Hi Ventana-users,

we have a special technical problem with our Ultra. It seems, that the liquid in the waste-tube is sometime pressed out of the hole on the platform like a fountain.

You can imagine the nice surprise, when the oil drops from the Ultra-roof onto the carousel-top. Everything is oily.

The technical service has'nt found a cause until now. The filter in the tube has been changed.

Has anyone seen a similar problem? Solved the problem?



Gudrun Lang



------------------------------

Message: 3
Date: Wed, 2 Oct 2013 13:47:22 -0400
From: Kim Donadio <one_angel_secret <@t> yahoo.com>
Subject: Re: [Histonet] RE: MOHS IPs
To: "Bauer, Karen L." <Bauer.Karen <@t> mayo.edu>
Cc: "histonet <@t> lists.utsouthwestern.edu"
        <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <628C0E60-8664-4DBC-B558-F3B5D9FEABF8 <@t> yahoo.com>
Content-Type: text/plain;       charset=us-ascii

What detection kit are you using?

Sent from my iPhone

On Oct 2, 2013, at 10:47 AM, "Bauer, Karen L." <Bauer.Karen <@t> mayo.edu> wrote:

> Thanks for the reply,
>
> We are pre-fixing in a formalin substitute, but we'll give the acetone a try.  I have diluted the protease, but have not gotten any good results.  We'll keep trying with that as well.
>
> We've tried multiple incubation times for the cytokeratin... From 5 minutes all the way up to 30 minutes... With no luck.
>
> We are using an enhanced polymer DAB.
>
> Thanks for the suggestions!  I greatly appreciate it!  :)
>
> Karen
>
> Karen L. Bauer HTL/HT (ASCP) | Histology Supervisor | Pathology | MOHS
> Lab Supervisor | Dermatology | Phone: 715-838-3205 |
> bauer.karen <@t> mayo.edu | Mayo Clinic Health System | 1221 Whipple Street
> | Eau Claire, WI 54702 | mayoclinichealthsystem.org
>
>
>
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
> pruegg <@t> ihctech.net
> Sent: Wednesday, October 02, 2013 9:36 AM
> To: Barbara Tibbs; Bauer, Karen L.; 'histonet <@t> lists.utsouthwestern.edu'
> Subject: RE: [Histonet] RE: MOHS IPs
>
>
>   I  agree with Barbara try fixing in cold acetone/ethanol mix even =
>   just  for  a  couple  of minutes then go directly to buffer do not dry
>   after=x,  I  would dilute the protease 1:4 with buffer (PBS or TBS
>   what  ever  u  u=) and do it for a short time.  Enzyme digestion on
>   frozen  sections  i=ricky  but  fixing a little and diluting the
>   enzyme  should  help  prevent  ov=  digestion.   How  long  are  u
>   incubating  the  primary?   Are  u usin=n enhanced labeled polymer
>   detection?  HRP/Dab or AEC or Alk.P/fast=d?
>
>
>
>
>
>
>
>
> -------- Original Message --------
>
> Subject: [Histonet] RE: MOHS =s
>
> From: Barbara Tibbs <[1]barbara.tibbs <@t> accuratediagnosticlabs.com>
>
> D=e: Wed, October 02, 2013 5:54 am
>
> To: "Bauer, Karen L." <[2]Bauer.Karen <@t> mayo.edu>,
>
> "'histonet <@t> lists.utsouthwest=n.edu'"
>
> <[3]histonet <@t> lists.utsouthwestern.edu>
>
>
>
> Hello Karen,
>   =A
>
> Back  in the old days of doing IHC manually on mostly fresh, froze   n  tissue  I would immediately place the slide with the frozen section
>   into  a=plin  jar  of  acetone  to  fix it. Keep the coplin jar of
>   acetone  in  the  cr=stat.  Leave  it  in  the acetone for only 10
>   minutes.  Try diluting the Pr=ease 2 - 1:1 to make it gentler so it
>   doesn't  "eat"  the  tissue.  This  te=nique worked well when I was
>   doing ER/PR on frozen breast tumors. Not su= it will work with skin
>   but it's worth a try.
>
>
>
> Barbara S. Tib=
>
> Histology Supervisor
>
> Accurate Diagnostic Labs
>
> South Pl=nfield, NJ
>
> [4]barbara.tibbs <@t> accuratediagnosticlabs.com
>
> 732-839-3374
>
> C=l: 610-809-6508
>
>
>
>
>
> _____________________________________=_
>
> From:                  [5]histonet-bounces <@t> lists.utsouthwestern.edu
>   <[6]histonet-bounces <@t> lists.utsouthwestern=du>  on behalf of Bauer,
>   Karen L. <[7]Bauer.Karen <@t> mayo.edu>
>
> Sent: Tuesday, October 01, 20 5:22 PM
>
> To: '[8]=stonet <@t> lists.utsouthwestern.edu'
>
> Subject: [Histonet] MOHS IPs
>
>
> Hi all,
>
>
>
> We  are  in the process of validating some a=ibodies in our MOHS
>   lab.  The Melan A (Mart 1) antibody is working well, =t it could be
>   darker.  We will be increasing the Ab incubation time to se=f that
>   will help.
>
>
>
> As  for  the Pan Keratin, we cannot get it = work at all. We use
>   Protease  2  on  our Ultra stainer for FFPE tissues in=stology, but
>   this  stain in MOHS is placed on fresh, unfixed tissue... by=anual
>   drop  method.  Any  time we've tried to use an enzyme retrieval, th=
>   tissue looks eaten up... even if we incubate if for a minute.
>
>
>   =ASince  the tissue is not fixed, I figured that no retrieval step
>   would  be=eded, but the Pan Keratin refuses to work with or without
>   retrieval.
>   =A
>
> For  those of you in MOHS labs that are using a manual staining me   thod  for  Melan A and Pan Keratin, would you be willing to share your
>   protoc= with us?
>
>
>
> Thanks so much!!
>
>
>
> Karen
>
>
>
> K=en L. Bauer HTL/HT (ASCP) | Histology Supervisor | Pathology |
>   MOHS   Lab   S=ervisor  |  Dermatology  |  Phone:  715-838-3205  |
>   [9]bauer.karen <@t> mayo.edu<[10]mailto:bauer.karen <@t> mayo.edu> | Mayo Clinic
>   Health  System  |  122=hipple  Street  |  Eau Claire, WI 54702 |
>   [11]mayoclinichealthsystem.org<http://www.[12]mayoclinichealthsystem.o
>   rg/>
>
>
>   =A
>
> _______________________________________________
>
> Histonet ma=ing list
>
> [13]Histo=t <@t> lists.utsouthwestern.edu
>
> [14]http://lists.utsouthwestern.edu/mailman/l=tinfo/histonet
>
>
>
> _________________________________________=____
>
> Histonet mailing list
>
> [15]Histonet <@t> lists.utsouthwestern.edu
>
> [16]http://lists.utso=hwestern.edu/mailman/listinfo/histonet
>
>
>
>
>
>
>
> References
>
>   1. 3D"mailto:barbara.tibbs <@t> accurated   2. ="mailto:Bauer.Karen <@t> mayo.edu"
>   3. 3D"mailto:histonet <@t> lists.utsouthwestern.edu   4. 3D"mailto:barbara.tibbs <@t> accuratediagnosticlabs.c   5. 3D"mailto:histonet-bounces <@t> lists.utsouthwestern.edu   6. 3D"mailto:histo   7. 3D"mailto:Bauer.Karen   8. 3D"mailto:histonet <@t> lists.utsouthwestern.edu"
>   9. 3D"mailto:bauer.kar  10. 3D"mailto:bauer.karen <@t> mayo  11.
> 3D"http://mayoclinichealt=  12. 3D"http:/  13. 3D"mailto:Histonet <@t> lists.utsouthwestern.edu"
>  14. 3D"http://lists.utsouthweste=
>  15. 3D"mailto:Histonet <@t> lists.u  16.
> file://localhost/tmp/3D"h_____________________________________________
> __
> Histonet mailing list
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------------------------------

Message: 4
Date: Wed, 2 Oct 2013 13:51:59 -0400
From: Kim Donadio <one_angel_secret <@t> yahoo.com>
Subject: Re: [Histonet] IHC turnaround time
To: "McKenzie, Emily" <McKenzie.Emily <@t> mhsil.com>
Cc: "histonet <@t> lists.utsouthwestern.edu"
        <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <75CAA15B-C45D-443E-87E9-E76143144E69 <@t> yahoo.com>
Content-Type: text/plain;       charset=utf-8

In less than 8 hours I can get 96 Ihc stains done with our dako link and that is two runs

Sent from my iPhone

On Oct 1, 2013, at 1:15 PM, "McKenzie, Emily" <McKenzie.Emily <@t> mhsil.com> wrote:

> Hello all,
> I am currently working on a Six Sigma Green Belt project and am looking for some data. Can any one give me their  general turnaround times for IHC, from time of order to time of delivery?
> Thanks for all your help with this,
>
> Emily K. McKenzie BS, HT(ASCP)
>
> Memorial Medical Center???701 North First Street???Springfield, IL
> 62781
> Ph: 217-788-3991???email: McKenzie.Emily <@t> mhsil.com
>
>
>
>
>  ________________________________
> This message (including any attachments) contains confidential information intended for a specific individual and purpose, and is protected by law. If you are not the intended recipient, you should delete this message. Any disclosure, copying, or distribution of this message, or the taking of any action based on it, is strictly prohibited.
> _______________________________________________
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> http://lists.utsouthwestern.edu/mailman/listinfo/histonet



------------------------------

Message: 5
Date: Wed, 2 Oct 2013 11:15:17 -0700
From: Jennifer MacDonald <JMacDonald <@t> mtsac.edu>
Subject: Re: [Histonet] Gram stain
To: Mesru T <turkekul <@t> gmail.com>
Cc: histonet <@t> lists.utsouthwestern.edu,
        histonet-bounces <@t> lists.utsouthwestern.edu
Message-ID:
        <OFFA2331AD.0B440E23-ON88257BF8.00644160-88257BF8.00644700 <@t> mtsac.edu>
Content-Type: text/plain; charset="US-ASCII"

We have had great success with the Twort gram stain.



From:   Mesru T <turkekul <@t> gmail.com>
To:     histonet <@t> lists.utsouthwestern.edu
Date:   10/02/2013 10:13 AM
Subject:        [Histonet] Gram stain
Sent by:        histonet-bounces <@t> lists.utsouthwestern.edu



Hi All,



Does anybody have a protocol for Gram stain on FFPE mouse intestine sections. We are looking to distinguish between Klebsiella and Vancomycin Resistant Enterococcus (VRE). I would like to avoid using Picric acid as some protocols suggest.

Thanks in advance.
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



------------------------------

Message: 6
Date: Wed, 02 Oct 2013 14:39:26 -0400
From: "Helene Degan" <deganh <@t> upstate.edu>
Subject: [Histonet] Hi
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <524C301E0200007A000271AE <@t> gatedom1.upstate.edu>
Content-Type: text/plain; charset=US-ASCII

Hi
Im looking for an acid phosphatase enzyme procedure that might be out there the one we tried in the past was difficult to work up and get good reproducible results.
Thanks

Helene D
Upstate University Medical
Syracuse, NY


------------------------------

Message: 7
Date: Wed, 2 Oct 2013 18:40:31 +0000
From: "Eytalis, Robert A" <Robert-Eytalis <@t> RiversideHealthCare.net>
Subject: [Histonet] RE: Validating new Benchmark instrument
To: Fawn Bomar <Fawn.Bomar <@t> HalifaxRegional.com>,
        "histonet <@t> lists.utsouthwestern.edu"
        <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
        <F3024E541CBF9049BD4A805E1646204A3C0EC820 <@t> RHCMBX01.RiversideHealthCare.net>

Content-Type: text/plain; charset="us-ascii"

I am thinking there has to be a way to integrate the Validation into the Optimization the vendor does. I think you should be able to make microarray blocks/slides of at least 10 positive and more for ERPR per CAP during the optimization. Now, the vendor will say they only do Optimization, but there this is a way to get  them to do the stains for you at least. You can create your own document for the Validation.
When your manager negotiates the cost of the analyzer, see if the vendor can help out with the initial expense with an allowance. It sounds like you might be past this point.
Has anyone else done it this way?

Robert A. Eytalis
Laboratory Manager
robert-eytalis <@t> riversidehealthcare.net
Phone: (815) 935-7256 ext. 5186
                (815) 935-7535
Fax         (815) 935-7068

Riverside Medical Center
350 N. Wall Street - Kankakee, IL 60901


http://mail.riversidehealthcare.net/owa/redir.aspx?C=qIzWpCGMNkq2SuiIt1v_2X7GmC1aOtAI7VlOiEQ974hhCQzwyDH6yJklmjoBIK92OWDFwivhIZs.&URL=http%3a%2f%2fwww.riversidemc.net%2f  |  http://mail.riversidehealthcare.net/owa/redir.aspx?C=qIzWpCGMNkq2SuiIt1v_2X7GmC1aOtAI7VlOiEQ974hhCQzwyDH6yJklmjoBIK92OWDFwivhIZs.&URL=http%3a%2f%2fwww.facebook.com%2fRiversideMC


________________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] on behalf of Fawn Bomar [Fawn.Bomar <@t> HalifaxRegional.com]
Sent: Tuesday, October 01, 2013 12:06 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Validating new Benchmark instrument

Hi everyone,



I know that this subject has come up several times and I am sorry to bother everyone again but I was hoping to find some help.  We are upgrading our Benchmark Immuno machines to the Benchmark XT immuno machine at the end of October.  Does anyone have any suggestions on how to approach the validation process?  Do we need to validate every antibody that we use as well as the kits?  I know that there is also a gray area surrounding how many slides need to be run to consider the antibody validated, but what would be the suggested amount of slides to validate a new machine as I know this will be very costly.  Does anybody have a log/checklist/spreadsheet that they would be willing to share to help document this validation of the machine and each of the antibodies?



Thank you in advance for your help



Fawn Bomar
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------------------------------

Message: 8
Date: Wed, 2 Oct 2013 18:51:22 +0000
From: Patricia F Lott <plott <@t> uab.edu>
Subject: [Histonet] need lever clamp for blade of leica microtome
To: "histonet <@t> lists.utsouthwestern.edu"
        <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
        <372C4AF089B6AE48B36F064FA891FF781380924D <@t> UABEXMB3.ad.uab.edu>
Content-Type: text/plain; charset="us-ascii"

Does anyone have an extra lever clamp for a Leica microtome?  I'd be happy to pay for it, and of course, for shipping!
Thanks,
Patty Lott
UAB CMBD Core Lab
205-934-2007


------------------------------

Message: 9
Date: Wed, 2 Oct 2013 14:03:38 -0500
From: Jan Shivers <shive003 <@t> umn.edu>
Subject: Re: [Histonet] Shipping to Countries Outside of the US
To: Sheila Miller <smmiller <@t> comhs.org>
Cc: "histonet <@t> lists.utsouthwestern.edu"
        <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
        <CAEoC1q3z_XnMMS5arjG6NTYeJE_8K35iWLWpewvmK1mRTWefvg <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

According to our Receiving department, each country/institution has its own
permit requirements for receiving samples from outside of their own
country.  It is up to the shipper to inquire of the receiver as to what
permits are needed to ship to that country (Germany, in your case).  My
source also said that FedEx would also have good information on this, since
they ship nearly everywhere around the globe.

Jan Shivers


On Wed, Oct 2, 2013 at 11:10 AM, Sheila Miller <smmiller <@t> comhs.org> wrote:

> Hello All,
>
> A question was posed to me by our Cancer Resource Department...
>
> I am wondering if you/lab has had experience with shipping tissue
> specimens outside the country.  We have a study where we may be doing this
> and they are asking if we have to have a special permit to ship outside our
> country.  I believe it is going to Germany.
>
> Thank you,
> Sheila
>
>
> ____________________________________
>
> This message and attachment(s), if any, is intended for the sole use of
> the individual and/or entity
> of which it is addressed, and may contain information that is
> privileged,confidential and prohibited
> from disclosure under applicable law. If you are not the addressee, or
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> behalf of the addressee, you are hereby notified that you may not use,
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> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>


------------------------------

Message: 10
Date: Wed, 2 Oct 2013 14:24:21 -0500
From: Yan Liang <yliang <@t> numirabio.com>
Subject: [Histonet] RE: Histotechnologist, (ASCP, QIHC certified) and
        Pathology assistant position in Seattle area
To: Brannon Owens <brannon <@t> alliedsearchpartners.com>,
        "histonet <@t> lists.utsouthwestern.edu"
        <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
        <8DC126BD0D840947A161BBF46AB31688035AE3305C <@t> DFW1MBX22.mex07a.mlsrvr.com>

Content-Type: text/plain; charset="iso-8859-1"


Needing a  Histotechnologist (ASCP, QIHC certified) or Scientist (Histopathology) for a Seattle area.  Five years of experience in routine histology including special stains, IHC and pathology knowledge.  Send an updated resume to yliang <@t> numirabio.com to be considered.  A full job description is on the Numirabio.com
..............................................
Yan Liang ?Ph.D.
Senior Director, Histopathology
Numira Biosciences Inc.
2023 120th Ave NE
Bellevue, WA 98005
Phone: 425-777-4950 ext 201
Cell: 425-830-3090
Fax:  425-777-4949
yliang <@t> numirabio.com
www.numirabio.com
www.altaportal.numirabio.com






------------------------------

Message: 11
Date: Wed, 2 Oct 2013 22:57:22 +0000
From: Teri Johnson <TJohnson <@t> gnf.org>
Subject: [Histonet] Vacuum and pressure in tissue processing
To: "histonet <@t> lists.utsouthwestern.edu"
        <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <9F3CFEE76E51B64991C7485270890B40497D12D6 <@t> EX5.lj.gnf.org>
Content-Type: text/plain; charset="us-ascii"

Dear friends,

I recall hearing at a conference (or maybe it was just a casual conversation by an expert during a NSH symposium break) that vacuum and pressure in tissue processing really accomplishes very little. I do believe that using heat and agitation of the solutions provides more activity kinetically and therefore makes processing more efficient.

Can someone affirm or deny the efficacy of vacuum and/or pressure in tissue processing, please?

Thank you, as always, for your wisdom.

Teri Johnson
Manager, Histology
Genomics Institute for
Novartis Research
Foundation
San Diego, CA
858-332-4752



------------------------------

Message: 12
Date: Wed, 2 Oct 2013 21:04:01 -0400
From: Gerald Davydov <geralddavydov <@t> gmail.com>
Subject: [Histonet] Trivew
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
        <CAO=5F_MBO-ezNR4pTryRUEPRce6Agmfm+1j5b9vpnYwhz4CEiw <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

Please share protocol for Triview from Zetta Corporation for Venatna.


------------------------------

Message: 13
Date: Thu, 3 Oct 2013 04:32:48 +0000
From: "Perow, Elliott S (PEROWES10)" <PEROWES10 <@t> juniata.edu>
Subject: [Histonet] Inconsistent Sections with Cryostat
To: "histonet <@t> lists.utsouthwestern.edu"
        <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
        <A1A3C2274BCBF94FA8A8E7B43C8830553EA9B43F <@t> Raphael.juniata.edu>
Content-Type: text/plain; charset="iso-8859-1"

Hello All,

I have a Leica Cryocut 1800 with a Reichert-Jung 2020 microtome that has sat for 8 years and went through a move from one academic building to another.  I am trying to section brook trout brain, but am having difficulty getting consistent sections.
I will get a good section and then the next section will be a very very thin shaving of usually just the top portion of the OCT block.  I have tried adjusting the clearance angle, made sure the blade and specimen is secured, replaced the blade with a new one, have tried different sectioning speeds, different thicknesses of the section and still continually only get a decent section every other turn.  I was wondering if anyone has had any similar experiences or if maybe the machine just needs a maintenance check due to it being an older machine.  I wasn't sure if the advancement mechanism may be off or if it is another issue that doesn't seem apparent to me.  Any help would be greatly appreciated.

Thanks,

Elliott Perow
Juniata College
Biology POE


CONFIDENTIALITY NOTICE: The materials in this electronic mail transmission (including all attachments) are private and confidential and are the property of the sender. The information contained in the material is privileged and is intended only for the use of the named addressee(s). If you are not the intended addressee, be advised that any unauthorized disclosure, copying, distribution or the taking of any action in reliance on the contents of this material is strictly prohibited. If you have received this e-mail in error, please immediately notify the sender by replying to the e-mail, and then destroy it immediately. Thank you.



------------------------------

Message: 14
Date: Thu, 3 Oct 2013 06:21:14 -0500
From: "Stephenson, Sheryl" <SStephenson <@t> lifecell.com>
Subject: [Histonet] HT HistoDeck question...
To: "histonet <@t> lists.utsouthwestern.edu"
        <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
        <C2A8FE4DA0ED8C468A3713F8607829D31DE32AE535 <@t> AMWPVEX03.kci.com>
Content-Type: text/plain; charset="us-ascii"

Hi,
Please clarify why this answer to the HistoDeck study question is  a) and not b).

Here is the question:

  'Frozen section slides cut from fresh, unfixed tissue specimens are optimally fixed in which of the following solutions?
a)       37%-40% formaldehyde
b)      Cold acetone
c)      Acetic acid alcohol
d)      Alcoholic formalin

Thanks,


Sheryl Stephenson | Histology Technician




_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



------------------------------

Message: 15
Date: Thu, 3 Oct 2013 07:54:56 -0400
From: "Manfre, Philip" <philip_manfre <@t> merck.com>
Subject: [Histonet] RE: Inconsistent Sections with Cryostat
To: "Perow, Elliott S (PEROWES10)" <PEROWES10 <@t> juniata.edu>,
        "histonet <@t> lists.utsouthwestern.edu"
        <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
        <558A4571351D0C42BD923F403F4198C4B27103726B <@t> USCTMXP51014.merck.com>
Content-Type: text/plain; charset="us-ascii"

I believe you may need to have the unit serviced.  It sounds like something is not tight enough, perhaps the stage or blade holder unit.  You said you secured everything which makes me think you have some issue with the cryostat itself.  If a sharp blade, tightened blade and specimen, and varying the speed doesn't work, then you may need professional assistance form a service technician, especially if it is an older unit that has been moved around.  Have you tried different temperatures?  I don't think that is the problem but it is worth a try.

Good luck! Cryosectioning can be an art in itself. Those decent sections are likely thicker than you intend, by the way.  Classic "thick and thin".


Philip Manfre, B.A., HT (ASCP)
Associate Principal Scientist
Merck Research Laboratories
WP45-251
PO Box 4
West Point, PA 19486

215-652-9750
215-993-0383 (fax)
philip_manfre <@t> merck.com




-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Perow, Elliott S (PEROWES10)
Sent: Thursday, October 03, 2013 12:33 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Inconsistent Sections with Cryostat

Hello All,

I have a Leica Cryocut 1800 with a Reichert-Jung 2020 microtome that has sat for 8 years and went through a move from one academic building to another.  I am trying to section brook trout brain, but am having difficulty getting consistent sections.
I will get a good section and then the next section will be a very very thin shaving of usually just the top portion of the OCT block.  I have tried adjusting the clearance angle, made sure the blade and specimen is secured, replaced the blade with a new one, have tried different sectioning speeds, different thicknesses of the section and still continually only get a decent section every other turn.  I was wondering if anyone has had any similar experiences or if maybe the machine just needs a maintenance check due to it being an older machine.  I wasn't sure if the advancement mechanism may be off or if it is another issue that doesn't seem apparent to me.  Any help would be greatly appreciated.

Thanks,

Elliott Perow
Juniata College
Biology POE


CONFIDENTIALITY NOTICE: The materials in this electronic mail transmission (including all attachments) are private and confidential and are the property of the sender. The information contained in the material is privileged and is intended only for the use of the named addressee(s). If you are not the intended addressee, be advised that any unauthorized disclosure, copying, distribution or the taking of any action in reliance on the contents of this material is strictly prohibited. If you have received this e-mail in error, please immediately notify the sender by replying to the e-mail, and then destroy it immediately. Thank you.

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------------------------------

Message: 16
Date: Thu, 3 Oct 2013 08:15:56 -0400
From: "Lee & Peggy Wenk" <lpwenk <@t> sbcglobal.net>
Subject: Re: [Histonet] HT HistoDeck question...
To: "Stephenson, Sheryl" <SStephenson <@t> lifecell.com>,
        <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <8FAB0141C65B42098CE4F110ECE9F372 <@t> HP2010>
Content-Type: text/plain; format=flowed; charset="iso-8859-1";
        reply-type=original

Personally, I think it's "a" is a wrong answer, and that you are correct
that "b" is a better answer. My students and I have found a couple of other
questions that we thought had the wrong answer indicated in the study set.

Peggy A. Wenk, HTL(ASCP)SLS
-----Original Message-----
From: Stephenson, Sheryl
Sent: Thursday, October 03, 2013 7:21 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] HT HistoDeck question...

Hi,
Please clarify why this answer to the HistoDeck study question is  a) and
not b).

Here is the question:

  'Frozen section slides cut from fresh, unfixed tissue specimens are
optimally fixed in which of the following solutions?
a) 37%-40% formaldehyde
b) Cold acetone
c) Acetic acid alcohol
d) Alcoholic formalin

Thanks,


Sheryl Stephenson | Histology Technician




_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

_______________________________________________
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Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet




------------------------------

Message: 17
Date: Thu, 3 Oct 2013 08:21:55 -0400
From: "Lee & Peggy Wenk" <lpwenk <@t> sbcglobal.net>
Subject: Re: [Histonet] RE: Inconsistent Sections with Cryostat
To: "Manfre, Philip" <philip_manfre <@t> merck.com>, "Perow, Elliott S
        \(PEROWES10\)" <PEROWES10 <@t> juniata.edu>,
        <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <F526A5B0D05046AA963690B931031FE1 <@t> HP2010>
Content-Type: text/plain; format=flowed; charset="iso-8859-1";
        reply-type=original

I just attended Jan Minshew's workshop on cryostats at the NSH Symposium in
Providence, RI, and she brought up something I had never thought of that
causes thick and thin.

If the handle that tightens the blade in the blade holder is over-tightened,
the blade will become bowed, and that will cause thick-and-thin during
sectioning.

According to Jan, the handle should be tightened to be parallel to the angle
of the blade holder "slope". A lot of times, the handle can be pushed
further back towards the back of the cryostat, beyond being parallel with
the slope. The thought seems to be, if tight is good, then tighter is
better, but not in this case.

Peggy A. Wenk, HTL(ASCP)SLS

-----Original Message-----
From: Manfre, Philip
Sent: Thursday, October 03, 2013 7:54 AM
To: Perow, Elliott S (PEROWES10) ; histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] RE: Inconsistent Sections with Cryostat

I believe you may need to have the unit serviced.  It sounds like something
is not tight enough, perhaps the stage or blade holder unit.  You said you
secured everything which makes me think you have some issue with the
cryostat itself.  If a sharp blade, tightened blade and specimen, and
varying the speed doesn't work, then you may need professional assistance
form a service technician, especially if it is an older unit that has been
moved around.  Have you tried different temperatures?  I don't think that is
the problem but it is worth a try.

Good luck! Cryosectioning can be an art in itself. Those decent sections are
likely thicker than you intend, by the way.  Classic "thick and thin".


Philip Manfre, B.A., HT (ASCP)
Associate Principal Scientist
Merck Research Laboratories
WP45-251
PO Box 4
West Point, PA 19486

215-652-9750
215-993-0383 (fax)
philip_manfre <@t> merck.com




-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Perow,
Elliott S (PEROWES10)
Sent: Thursday, October 03, 2013 12:33 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Inconsistent Sections with Cryostat

Hello All,

I have a Leica Cryocut 1800 with a Reichert-Jung 2020 microtome that has sat
for 8 years and went through a move from one academic building to another.
I am trying to section brook trout brain, but am having difficulty getting
consistent sections.
I will get a good section and then the next section will be a very very thin
shaving of usually just the top portion of the OCT block.  I have tried
adjusting the clearance angle, made sure the blade and specimen is secured,
replaced the blade with a new one, have tried different sectioning speeds,
different thicknesses of the section and still continually only get a decent
section every other turn.  I was wondering if anyone has had any similar
experiences or if maybe the machine just needs a maintenance check due to it
being an older machine.  I wasn't sure if the advancement mechanism may be
off or if it is another issue that doesn't seem apparent to me.  Any help
would be greatly appreciated.

Thanks,

Elliott Perow
Juniata College
Biology POE


CONFIDENTIALITY NOTICE: The materials in this electronic mail transmission
(including all attachments) are private and confidential and are the
property of the sender. The information contained in the material is
privileged and is intended only for the use of the named addressee(s). If
you are not the intended addressee, be advised that any unauthorized
disclosure, copying, distribution or the taking of any action in reliance on
the contents of this material is strictly prohibited. If you have received
this e-mail in error, please immediately notify the sender by replying to
the e-mail, and then destroy it immediately. Thank you.

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Notice:  This e-mail message, together with any attachments, contains
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not the intended recipient, and have received this message in error,
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------------------------------

Message: 18
Date: Thu, 3 Oct 2013 09:40:06 -0400
From: "Watson, Linda" <Linda.Watson <@t> bms.com>
Subject: RE: [Histonet] HT HistoDeck question...
To: Lee & Peggy Wenk <lpwenk <@t> sbcglobal.net>, "Stephenson, Sheryl"
        <SStephenson <@t> lifecell.com>, "histonet <@t> lists.utsouthwestern.edu"
        <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
        <351A66CE7FB11D40AB8AC8FE5559EC7E11A975A411 <@t> ushpwbmsmmp008.one.ads.bms.com>

Content-Type: text/plain; charset="us-ascii"

For frozen cut sections, would the fixation also depend on what you plan on doing with it. For example, H&E, Special Stain or IHC? Please correct if I am wrong. I think that is a trick question!!!

>-----Original Message-----
>From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-
>bounces <@t> lists.utsouthwestern.edu] On Behalf Of Lee & Peggy Wenk
>Sent: Thursday, October 03, 2013 8:16 AM
>To: Stephenson, Sheryl; histonet <@t> lists.utsouthwestern.edu
>Subject: Re: [Histonet] HT HistoDeck question...
>
>Personally, I think it's "a" is a wrong answer, and that you are correct
>that "b" is a better answer. My students and I have found a couple of
>other
>questions that we thought had the wrong answer indicated in the study
>set.
>
>Peggy A. Wenk, HTL(ASCP)SLS
>-----Original Message-----
>From: Stephenson, Sheryl
>Sent: Thursday, October 03, 2013 7:21 AM
>To: histonet <@t> lists.utsouthwestern.edu
>Subject: [Histonet] HT HistoDeck question...
>
>Hi,
>Please clarify why this answer to the HistoDeck study question is  a)
>and
>not b).
>
>Here is the question:
>
>  'Frozen section slides cut from fresh, unfixed tissue specimens are
>optimally fixed in which of the following solutions?
>a) 37%-40% formaldehyde
>b) Cold acetone
>c) Acetic acid alcohol
>d) Alcoholic formalin
>
>Thanks,
>
>
>Sheryl Stephenson | Histology Technician
>
>
>
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet

This message (including any attachments) may contain confidential, proprietary, privileged and/or private information.  The information is intended to be for the use of the individual or entity designated above.  If you are not the intended recipient of this message, please notify the sender immediately, and delete the message and any attachments.  Any disclosure, reproduction, distribution or other use of this message or any attachments by an individual or entity other than the intended recipient is prohibited.



------------------------------

Message: 19
Date: Thu, 3 Oct 2013 13:42:34 +0000
From: Bernice Frederick <b-frederick <@t> northwestern.edu>
Subject: RE: [Histonet] HT HistoDeck question...
To: "Watson, Linda" <Linda.Watson <@t> bms.com>, Lee & Peggy Wenk
        <lpwenk <@t> sbcglobal.net>, "Stephenson, Sheryl"
        <SStephenson <@t> lifecell.com>,     "histonet <@t> lists.utsouthwestern.edu"
        <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
        <62C639732D3F274DACED033EBDF6ADAF2F00D4EA <@t> evcspmbx2.ads.northwestern.edu>

Content-Type: text/plain; charset="us-ascii"

We fix H&E's in 95%  and our IHC protocol is acetone/alcohol fixation.

Bernice Frederick HTL (ASCP)
Senior Research Tech
Pathology Core Facility
ECOGPCO-RL
Robert. H. Lurie Cancer Center
Northwestern University
710 N Fairbanks Court
Olson 8-421
Chicago,IL 60611
312-503-3723
b-frederick <@t> northwestern.edu

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Watson, Linda
Sent: Thursday, October 03, 2013 8:40 AM
To: Lee & Peggy Wenk; Stephenson, Sheryl; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] HT HistoDeck question...

For frozen cut sections, would the fixation also depend on what you plan on doing with it. For example, H&E, Special Stain or IHC? Please correct if I am wrong. I think that is a trick question!!!

>-----Original Message-----
>From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-
>bounces <@t> lists.utsouthwestern.edu] On Behalf Of Lee & Peggy Wenk
>Sent: Thursday, October 03, 2013 8:16 AM
>To: Stephenson, Sheryl; histonet <@t> lists.utsouthwestern.edu
>Subject: Re: [Histonet] HT HistoDeck question...
>
>Personally, I think it's "a" is a wrong answer, and that you are
>correct that "b" is a better answer. My students and I have found a
>couple of other questions that we thought had the wrong answer
>indicated in the study set.
>
>Peggy A. Wenk, HTL(ASCP)SLS
>-----Original Message-----
>From: Stephenson, Sheryl
>Sent: Thursday, October 03, 2013 7:21 AM
>To: histonet <@t> lists.utsouthwestern.edu
>Subject: [Histonet] HT HistoDeck question...
>
>Hi,
>Please clarify why this answer to the HistoDeck study question is  a)
>and not b).
>
>Here is the question:
>
>  'Frozen section slides cut from fresh, unfixed tissue specimens are
>optimally fixed in which of the following solutions?
>a) 37%-40% formaldehyde
>b) Cold acetone
>c) Acetic acid alcohol
>d) Alcoholic formalin
>
>Thanks,
>
>
>Sheryl Stephenson | Histology Technician
>
>
>
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet

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------------------------------

Message: 20
Date: Thu, 3 Oct 2013 09:59:58 -0400
From: "Pam Barker" <relia1 <@t> earthlink.net>
Subject: [Histonet] The NSH was AMAZING!!! Congratulations to the 2013
        Leadership,     Education and Advocacy Award Winners!! From Pam Barker and
        RELIA   Solutions!!
To: "Histonet" <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <006001cec040$d9e6e1f0$8db4a5d0$@earthlink.net>
Content-Type: text/plain;       charset="us-ascii"

Hi Histonetters!!!
How are you doing?
The NSH was AMAZING!!!!!
Providence was GORGEOUS!!!!
The Speakers were BRILLIANT!!!!

CONGRATULATIONS TO THE 2013 Leadership Education And Advocacy Award Winners:
Histotechnologist of the Year-Wanda Jones
J.B. McCormick Award- Peggy Wenk
President's Award- Janet Dapson
Lee G. Luna Foreign Travel Scholarship- M. Lamar Jones
William Hacker Memorial Award- Anthony Wong
Rosemary & Donald Ostermeier Memorial Award- Christiane Coady
Ventana IHC Award- Tiana Baskin
Biogenex Standardization in IHC Award-David Davis
Biogenex Standardization in ISH Award Dale Telgenhoff
Dako Standardization of IHC Award-Jennifer Wright
Anne Preece Award -Sarah Mack
Leica Leadership in Management Award-Nirmala Amin
Leica Leadership in Teaching Award- Elaine Basham
Newcomer Helping Hand Award- Jean Mitchell
Epitomics IHC Award- Melissa Downing
Jules Elias Excellence in IHC Award- James Burchette

Since I have returned to the office my phone has been ringing off the hook
with exciting new opportunities.  Here is a list of my current openings.
All of these are full time permanent positions.  My clients offer excellent
compensation, benefits and relocation assistance.
And they are ready to interview and hire right away!

HISTOLOGY SUPERVISORS/MANAGERS:
Histology Supervisor - Tallahassee, FL
Histology Supervisor - West Palm Beach, FL
Histology Supervisor - Seattle, WA
Lead Pathology Tech - East of Columbus, OH

HISTOTECHNICIANS/HISTOTECHNOLOGISTS:
Histology Tech with Electron Microscopy - Southern CA
Histotechnician Day shift -East of Columbus, OH
Histotechnician Day shift - Tyler, TX
Histotechnologist - Days - Seattle, WA
Histology Tech - West Palm Beach, FL
Histotechnician - Days - Harrisonburg, VA
Histotechnician - Nights - Nashville, TN
Senior Histotechnologist with FISH - Louisville, KY

OTHER OPPORTUNITIES
MT (ASCP) with Molecular experience- RTP, SC

Of course I can't put all the information about these opportunities in an
e-mail.  So if you or anyone you know might be interested in hearing more
about any of these positions or want help with a job search in another area
please contact me.  I can be reached at 866-607-3542 or
relia1 <@t> earthlink.net.

Remember it never hurts to look.  Thanks-Pam

Right Place, Right Time, Right Move with RELIA!

Thank You!
 Pam M. Barker

Pam Barker
President/Senior Recruiting Specialist-Histology
RELIA Solutions
Specialists in Allied Healthcare Recruiting
5703 Red Bug Lake Road #330
Winter Springs, FL 32708-4969
Phone: (407)657-2027
Cell:     (407)353-5070
FAX:     (407)678-2788
E-mail: relia1 <@t> earthlink.net
www.facebook.com/PamBarkerRELIA
www.linkedin.com/in/reliasolutions
www.twitter.com/pamatrelia










------------------------------

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End of Histonet Digest, Vol 119, Issue 4
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