[Histonet] Work hours reply to patti loykosek
Franssen, Dr. Catherine
cfranssen <@t> nsalabs.com
Wed May 23 14:22:53 CDT 2007
Patti- We're in the process of switching our histology staff to 5 8hr days from the 4 10hr days system. Originally it was implemented to account for long staining and slicing procedures that we specialize in, but improved technology makes that less of an issue and staggered times of arrival (8-4 or 9-5) take care of it. From the histologists' viewpoint the 4 10s were great because they could choose a Friday or Monday to take off and often they could take 4 day weekends, switching off weekends. (One or two part-timers cover any necessary weekend work, with full-timers available as needed.) It was great for morale and there was a nice coffee-and-doughnut kinship among the 7am crew (management/administration has always worked 5 8's... or 5 10s as it often turns into!). From the management perspective it got too difficult to work at half-staff for 2 days of the week. For a long time the part-timers filled in on those days, but as a small company, most of our histologists are senior and are very involved with project management... not having them in to supervise the part-time techs and communicate with clients pushed us over the edge. Plus, with increasing volumes, scheduling the work/specimen flow got too complex. Another note: on the 4-day system it is much more noticeable and hard to recover from someone taking a sick or personal day. In a larger company some of these issues may not exist... but there's my $0.04.
Hello to HistoLand.
Catherine
Catherine Lowry Franssen, Ph.D.
NeuroScience Associates
10915 Lake Ridge Drive
Knoxville, TN 37934
tel. 865-675-2245
cel. 865-712-9314
fax. 865-675-2787
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Sent: Wednesday, May 23, 2007 1:09 PM
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Subject: Histonet Digest, Vol 42, Issue 33
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Today's Topics:
1. RE: Histonet Digest, Vol 42, Issue 32 (Kobler, James)
2. Ethanol ready-to-use for LCM (Castillos, Luminita)
3. RE: NK cells in mouse tissue and PLP fixation (Gayle Callis)
4. RE: -20C acetone fixative (C.M. van der Loos)
5. RE: RE: -20C acetone fixative (Edwards, R.E.)
6. Cryomold adapters (Martha Ward)
7. Cell Blocks on small buttons... (Lott, Robert)
8. celloidin-ether/alc (Mary Lou Norman)
9. Work hours (Patti Loykasek)
10. RE: celloidin-ether/alc (Rittman, Barry R)
11. Re: Work hours (Rene J Buesa)
12. RE: Work hours (Mike Pence)
----------------------------------------------------------------------
Message: 1
Date: Wed, 23 May 2007 10:20:34 -0400
From: "Kobler, James" <JKOBLER <@t> PARTNERS.ORG>
Subject: [Histonet] RE: Histonet Digest, Vol 42, Issue 32
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<6D1DFC2837CEAE4BBBDED596071430854D07D4 <@t> PHSXMB4.partners.org>
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Dear histonet contributors,
I would greatly appreciate any advice about antibodies that work well with
ferret tissue (and any related processing tips). We are particularly interested
in antibodies to markers for fibroblasts, myofibroblasts, endothelial, smooth
muscle and inflammatory cells, as well as extracellular matrix. Thanks very
much,
Jim Kobler, Massachusetts General Hospital
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Message: 2
Date: Wed, 23 May 2007 10:28:19 -0400
From: "Castillos, Luminita" <lcastillos <@t> cogenics.com>
Subject: [Histonet] Ethanol ready-to-use for LCM
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<DE517009A0E1B345BAE029F86CADECCB059934 <@t> ncmsx2.gpi.genaissance.com>
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Hi,
Does any know from where to buy ready-to-use ethanol (100%, 95%, 70%)
molecular biology grade and RNase-free, compatible for Laser Capture
microdissection staining protocol?. Thank you very much.
L,
------------------------------
Message: 3
Date: Wed, 23 May 2007 08:43:03 -0600
From: Gayle Callis <gcallis <@t> montana.edu>
Subject: RE: [Histonet] NK cells in mouse tissue and PLP fixation
To: Bruce Abaloz <brucea <@t> unimelb.edu.au>,
Histonet <@t> lists.utsouthwestern.edu
Message-ID:
<6.0.0.22.1.20070523083057.01b3a148 <@t> gemini.msu.montana.edu>
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Sonya,
You can try PLP immersion fixation followed by sucrose cryoprotection, but
the publication I read (reference was in a Histonet message yesterday) did
a fixation study with PLP and other fixatives. They had little success
with the NK1.1 (P136) antibody following immersion fixation with all
fixatives, before snap freezing or after fresh tissue frozen sections were
cut. They had nice staining following PERFUSION (with PLP) of the
mouse. After perfusion, and once the tissues are dissected out, then
immersion into this fixative should ensure complete fixation. You may want
to do final fixation in PLP overnight, some do it for 5 - 6 hours longer
here, then do the 30% sucrose cryoprotection at 4C. Just remove the tissue
from fixative and go to the sucrose solution as Bruce suggested. Small
fixed tissues often float on top of the sucrose solution. A general rule
of thumb the that cryoprotection is done, tissues sink to the bottom of the
container.
Some people use a sucrose gradient, often seen in the literature, but we
have never found that necessary. We blot off the excess sucrose solution
before embedding in OCT.
Hopefully you have access to a perfusion methods IF your immersion fails.
Gayle Callis HTL, HT, MT(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717
At 06:12 PM 5/22/2007, you wrote:
>I just snap frooze the fresh tissue in isopentane on dry ice - this has
>worked with all the other markers I've been looking at but I gather NK
>cells are more tricky.
>I want to try PLP fixation before I freeze so do I put it straight from
>PLP to 30% sucrose or do I wash/use increasing amounts of sucrose.
>Once I've left the tissue overnight in sucrose can I then freeze it in
>TissueTek on dry ice as normal?
>
>Thanks
>Sonya
>
>-----Original Message-----
>From: Gayle Callis [mailto:gcallis <@t> montana.edu]
>Sent: 22 May 2007 15:49
>To: Martin S.
>Subject: Re: [Histonet] NK cells in mouse tissue and PLP fixation
>
>How did you fix in the first place?
>
>You need to sucrose cryoprotect after PLP fixation, in 30% sucrose
>overnight BEFORE snap freezing the tissues.
>
>At 04:01 AM 5/22/2007, you wrote:
>>Has anyone done any staining for NK cells in mouse tissue. We have an
>>antibody against NK1.1 (PK136) but it doesnt seem to work on
>>fresh-frozen tissues. I have looked through the literature and some
>>people seem to have got iot to work on fresh-frozens while others have
>>used PLP fixation before embedding in Tissue Tek.
>>
>>Has anyone had any experience with this Ab?
>>
>>Also, if I fix the tissues with PLP before freezing then how should I
>>treat them before outting them in TissueTek to freeze?
>>
>>Thanks
>>Sonya
>>
>>
>>
>>_______________________________________________
>>Histonet mailing list
>>Histonet <@t> lists.utsouthwestern.edu
>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>Gayle Callis
>MT,HT,HTL(ASCP)
>Research Histopathology Supervisor
>Veterinary Molecular Biology
>Montana State University - Bozeman
>PO Box 173610
>Bozeman MT 59717-3610
>
>
>
>
>
>
>
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>--
>BRUCE ABALOZ
>HISTOLOGIST PH:61383446282
>DEPT. of ZOOLOGY EMAIL: brucea <@t> unimelb.edu.au
>THE UNIVERSITY of MELBOURNE. FAX:61383447909
>VICTORIA. AUSTRALIA 3010
> Nobody Can Make You Feel Inferior Without YOUR Permission -
> Eleanor Roosevelt
> WHETHER YOU THINK YOU CAN-OR WHETHER YOU THINK YOU
> CAN'T-YOU'RE RIGHT!!
> Be reasonable... Demand the
> impossible.....
> <')))>>< <')))>><
> <')))>>< <')))>><
> P Please consider the environment before printing this e-mail.
>
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------------------------------
Message: 4
Date: Wed, 23 May 2007 16:48:47 +0200
From: "C.M. van der Loos" <c.m.vanderloos <@t> amc.uva.nl>
Subject: [Histonet] RE: -20C acetone fixative
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <37e62d383380.38338037e62d <@t> amc.uva.nl>
Content-Type: text/plain; charset="us-ascii"
Long time ago they told me it is a milder fixative at -20C than at RT
???
Chris van der Loos, PhD
Dept. of Pathology
Academic Medical Center M2-230
Meibergdreef 9
NL-1105 AZ Amsterdam
The Netherlands
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[[1]mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Charles,
Roger
Sent: Tuesday, May 22, 2007 12:29 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] -20C acetone fixative
Does any know or remember why acetone, when used as a fixative, is
always used "ice cold" or in our case at -20C?
Roger Charles
Microbiologist
Pennsylvania Veterinary Laboratory
2305 N Cameron St
Harrisburg, PA 17110
717-787-8808
References
1. javascript:main.compose('new', 't=histonet-bounces <@t> lists.utsouthwestern.edu')
------------------------------
Message: 5
Date: Wed, 23 May 2007 16:15:14 +0100
From: "Edwards, R.E." <ree3 <@t> leicester.ac.uk>
Subject: RE: [Histonet] RE: -20C acetone fixative
To: "C.M. van der Loos" <c.m.vanderloos <@t> amc.uva.nl>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<DC88BEDFD1FC3F468D0376A7C75465F7101C6E24 <@t> Saffron.cfs.le.ac.uk>
Content-Type: text/plain; charset="US-ASCII"
And me, in particular for enzyme histochemistry...
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of C.M. van
der Loos
Sent: 23 May 2007 15:49
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] RE: -20C acetone fixative
Long time ago they told me it is a milder fixative at -20C than at
RT
???
Chris van der Loos, PhD
Dept. of Pathology
Academic Medical Center M2-230
Meibergdreef 9
NL-1105 AZ Amsterdam
The Netherlands
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[[1]mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf
Of
Charles,
Roger
Sent: Tuesday, May 22, 2007 12:29 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] -20C acetone fixative
Does any know or remember why acetone, when used as a fixative, is
always used "ice cold" or in our case at -20C?
Roger Charles
Microbiologist
Pennsylvania Veterinary Laboratory
2305 N Cameron St
Harrisburg, PA 17110
717-787-8808
References
1. javascript:main.compose('new',
't=histonet-bounces <@t> lists.utsouthwestern.edu')
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------------------------------
Message: 6
Date: Wed, 23 May 2007 11:02:18 -0400
From: "Martha Ward" <mward <@t> wfubmc.edu>
Subject: [Histonet] Cryomold adapters
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<61135F0455D33347B5AAE209B903A3041B1C9EBD <@t> EXCHVS2.medctr.ad.wfubmc.edu>
Content-Type: text/plain; charset="us-ascii"
I am looking for cryomold adapters and I hope someone can help. The
ones we are using have a round center and are a white, hard plastic. I
can find adapters that have a square center but that isn't what we want.
I have tried the usual vendors but was hoping someone could help me.
Thanks in advance for your help.
Martha Ward, MT (ASCP) QIHC
Assistant Manager, Molecular Diagnostics Lab
Wake Forest University Baptist Medical Center
Medical Center Blvd.
Winston-Salem, NC 27157
336-716-2756
mward <@t> wfubmc.edu
------------------------------
Message: 7
Date: Wed, 23 May 2007 10:23:14 -0500
From: "Lott, Robert" <Robert.Lott <@t> TriadHospitals.com>
Subject: [Histonet] Cell Blocks on small buttons...
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<673832E27C45FC4D97EF758FFC777C27020E03DC <@t> CPRTEVS01.triadhospitals.net>
Content-Type: text/plain; charset="us-ascii"
Hi Everyone,
We are interested in hearing about different procedures for recovering
small cytology aspirates (any fluid really!) ... and making a cell block
from them using some type of coagulative process/fluid/etc. ... so that
virtually all of the cells are retained!
There may be some commercial products available ... would like to hear
about those as well.!!
Thanks!!!!
Robert
Robert L. Lott, HTL(ASCP)
Manager, Anatomic Pathology
LabFirst / Trinity Medical Center - formerly
Montclair Baptist Medical Center
800 Montclair Road
Birmingham, AL 35213
205-592-5388 phone
205-592-5646 fax
robert.lott <@t> triadhospitals.com
------------------------------
Message: 8
Date: Wed, 23 May 2007 11:26:13 -0400
From: Mary Lou Norman <mlm11 <@t> cornell.edu>
Subject: [Histonet] celloidin-ether/alc
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<6.2.1.2.2.20070523111029.04899930 <@t> postoffice9.mail.cornell.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed
Hello Histonet,
Is there any other choice besides the cellodin-ether/alcohol coating? My
parloidin is taking 'forever' to dissolve -- I have read a week but I could
have used it a week ago.
Thanks for your help.
Mary Lou Norman
------------------------------
Message: 9
Date: Wed, 23 May 2007 08:42:22 -0700
From: Patti Loykasek <ploykasek <@t> phenopath.com>
Subject: [Histonet] Work hours
To: histonet <histonet <@t> pathology.swmed.edu>
Message-ID: <C279AEEE.FD8D%ploykasek <@t> phenopath.com>
Content-Type: text/plain; charset="US-ASCII"
I was wondering if anyone in histoland has experienced moving from a
schedule of standard 8 hour day/5 days a week to a 10 hour day/4 days a
week with techs having a day off during the week. If so, what are the pros
and cons of this type of schedule. I'm interested in feedback from both
bench techs and supervisor/managers. We are in the initial stages of
contemplating this and would appreciate info from anyone that has tried it.
Thank you.
Patti Loykasek BS, HTL, QIHC
PhenoPath Laboratories
Seattle, WA
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and destroy all copies of the original message, or you may call PhenoPath
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------------------------------
Message: 10
Date: Wed, 23 May 2007 11:37:45 -0500
From: "Rittman, Barry R" <Barry.R.Rittman <@t> uth.tmc.edu>
Subject: RE: [Histonet] celloidin-ether/alc
To: "Mary Lou Norman" <mlm11 <@t> cornell.edu>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<EA1FDD2A141B7448B4B1AFFFCAC08DE406169F41 <@t> UTHEVS1.mail.uthouston.edu>
Content-Type: text/plain; charset="iso-8859-1"
Mary Lou
Will dissolve much more rapidly if soak the strips first in absolute ethanol for a day o so. The add the ether.
Barry
________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Mary Lou Norman
Sent: Wed 5/23/2007 10:26 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] celloidin-ether/alc
Hello Histonet,
Is there any other choice besides the cellodin-ether/alcohol coating? My
parloidin is taking 'forever' to dissolve -- I have read a week but I could
have used it a week ago.
Thanks for your help.
Mary Lou Norman
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 11
Date: Wed, 23 May 2007 09:52:11 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Work hours
To: Patti Loykasek <ploykasek <@t> phenopath.com>, histonet
<histonet <@t> pathology.swmed.edu>
Message-ID: <990345.12651.qm <@t> web61216.mail.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
I have tried it.
Cons: If yu don't have a flow of specimens in 4 days equivalent to the flow on 5 days, the personnel will be idling some time each day. If specimens or workload is available everybody will have work to do.
Pros: it is fantastic having 3 days off weekly. Everybody likes that!
René J.
Patti Loykasek <ploykasek <@t> phenopath.com> wrote:
I was wondering if anyone in histoland has experienced moving from a
schedule of standard 8 hour day/5 days a week to a 10 hour day/4 days a
week with techs having a day off during the week. If so, what are the pros
and cons of this type of schedule. I'm interested in feedback from both
bench techs and supervisor/managers. We are in the initial stages of
contemplating this and would appreciate info from anyone that has tried it.
Thank you.
Patti Loykasek BS, HTL, QIHC
PhenoPath Laboratories
Seattle, WA
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and destroy all copies of the original message, or you may call PhenoPath
Laboratories, Seattle, WA U.S.A. at (206) 374-9000.
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Message: 12
Date: Wed, 23 May 2007 11:58:01 -0500
From: "Mike Pence" <mpence <@t> grhs.net>
Subject: RE: [Histonet] Work hours
To: "Rene J Buesa" <rjbuesa <@t> yahoo.com>, "Patti Loykasek"
<ploykasek <@t> phenopath.com>, "histonet" <histonet <@t> pathology.swmed.edu>
Message-ID: <661949901A768E4F9CC16D8AF8F2838CA1C5E3 <@t> IS-E2K3.grhs.net>
Content-Type: text/plain; charset="iso-8859-1"
How do you decide who gets which days off! Are they floating days?
Mike
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Wednesday, May 23, 2007 11:52 AM
To: Patti Loykasek; histonet
Subject: Re: [Histonet] Work hours
I have tried it.
Cons: If yu don't have a flow of specimens in 4 days equivalent to the flow on 5 days, the personnel will be idling some time each day. If specimens or workload is available everybody will have work to do.
Pros: it is fantastic having 3 days off weekly. Everybody likes that!
René J.
Patti Loykasek <ploykasek <@t> phenopath.com> wrote:
I was wondering if anyone in histoland has experienced moving from a schedule of standard 8 hour day/5 days a week to a 10 hour day/4 days a week with techs having a day off during the week. If so, what are the pros and cons of this type of schedule. I'm interested in feedback from both bench techs and supervisor/managers. We are in the initial stages of contemplating this and would appreciate info from anyone that has tried it. Thank you.
Patti Loykasek BS, HTL, QIHC
PhenoPath Laboratories
Seattle, WA
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you are not the intended recipient, please contact the sender by e-mail
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Laboratories, Seattle, WA U.S.A. at (206) 374-9000.
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