[Histonet] trap fixation and decal my 2 cents
Madary, Joseph
MadaryJ <@t> MedImmune.com
Thu Feb 4 09:45:39 CST 2010
The one thing I do recall was to cold decal in EDTA after cold fixation. I think it helped avoid some inconsistent staining, at least in our hands.
Nick Madary, HT/HTL(ASCP)QIHC
Medimmune Histology Mgr,
OMW, Area 4, Lab 2438
301.398.4745(vm)
301.398.6360(lab)
301.398.9745(fax)
-----Original Message-----
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Sent: Thursday, February 04, 2010 10:27 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 75, Issue 5
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Today's Topics:
1. GI Biopsies (Anderson, David W)
2. P63 (Sharon.Davis-Devine)
3. RE: P63 (Mike Pence)
4. RE: P63 (Drew Sally A)
5. RE: GI Biopsies (Mahoney,Janice A)
6. RE: Eosin leaching out of sections (Joyce Cline)
7. RE: Eosin leaching out of sections (Rathborne, Toni)
8. P63 RUO (Sharon.Davis-Devine)
9. RE: P63 RUO (McMahon, Loralee A)
10. Microarray info thanks! (mtitford <@t> aol.com)
11. Drying ovens, eosin fading, microtome PM, eyeballs
(Jeffrey Silverman)
12. Re: TRAP assay - acid phosphatase (Sherwood, Margaret )
13. slide drying (kyle ayres)
14. RE: TRAP assay - acid phosphatase (Liz Chlipala)
15. FW: FFPE Microwave processed biopsies (Delaney, Sondra L)
16. autofluorescence experiment (Nicole Collette)
17. Re: Derm Lab (soofia siddiqui)
18. Frozen Sections Slides per Day Question (soofia siddiqui)
19. mailers (Mary Lloyd)
20. RE: Tape/Film Coverslips vs. Glass (Percival Karen)
21. Re: Eosin leaching out of sections (rgrow <@t> bmnet.com)
22. Re: TRAP Assay and IHC (Sherwood, Margaret )
23. RE: Tape/Film Coverslips vs. Glass (Rosa Fields)
----------------------------------------------------------------------
Message: 1
Date: Wed, 3 Feb 2010 12:10:24 -0600
From: "Anderson, David W" <david.w.anderson <@t> Vanderbilt.Edu>
Subject: [Histonet] GI Biopsies
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<47299A40AEED044DA3255843A948CBCD03251A6F5A <@t> ITS-HCWNEM02.ds.Vanderbilt.edu>
Content-Type: text/plain; charset="us-ascii"
We are having an issue with our GI Biopsy H&E Stains. On our GI Biopsies, we currently cut eight slides, staining levels 2, 5 and 8 with H&E. The pathologist are noticing that the level two slide looks great but then levels 5 and 8 will have what they call "Blue Blobs" on the tissue sections. They tend to look like formalin salt crystals. Does anyone else experience this? Do any of you have a resolution to this problem? Thank you so much for your help with this matter.
David Anderson,HT(ASCP)
------------------------------
Message: 2
Date: Wed, 3 Feb 2010 12:13:09 -0600
From: "Sharon.Davis-Devine" <Sharon.Davis-Devine <@t> carle.com>
Subject: [Histonet] P63
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<50003EC02E2CEA4583BEB3CD08EAC1E090DDEE <@t> EXCHANGEBE2.carle.com>
Content-Type: text/plain; charset="us-ascii"
Do any of you Ventana users have a P63 that is not an RUO? If so, where
do you get it from? Thanks a bunch.
Sharon Davis-Devine, CT (ASCP)
Cytology-Histology Supervisor
Carle Foundation Hospital
Laboratory and Pathology Services
611 West Park Street
Urbana, Illinois 61801
217-383-3572
sharon.davis-devine <@t> carle.com
------------------------------
Message: 3
Date: Wed, 3 Feb 2010 12:20:52 -0600
From: "Mike Pence" <mpence <@t> grhs.net>
Subject: RE: [Histonet] P63
To: "Sharon.Davis-Devine" <Sharon.Davis-Devine <@t> carle.com>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<661949901A768E4F9CC16D8AF8F2838C017A3D37 <@t> is-e2k3.grhs.net>
Content-Type: text/plain; charset="us-ascii"
>From Cell Marque
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Sharon.Davis-Devine
Sent: Wednesday, February 03, 2010 12:13 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] P63
Do any of you Ventana users have a P63 that is not an RUO? If so, where
do you get it from? Thanks a bunch.
Sharon Davis-Devine, CT (ASCP)
Cytology-Histology Supervisor
Carle Foundation Hospital
Laboratory and Pathology Services
611 West Park Street
Urbana, Illinois 61801
217-383-3572
sharon.davis-devine <@t> carle.com
_______________________________________________
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 4
Date: Wed, 3 Feb 2010 12:21:56 -0600
From: "Drew Sally A" <SDrew <@t> uwhealth.org>
Subject: RE: [Histonet] P63
To: "Histonet" <histonet <@t> pathology.swmed.edu>
Message-ID:
<738A7878143FF74BB77436E255743C1A0100049E <@t> UWHC-MAIL03.uwhis.hosp.wisc.edu>
Content-Type: text/plain; charset="US-ASCII"
We use an IVD p63 (cat# CM163) from BioCare Medical. Contact me if
you'd like further info.
Sally Ann Drew, MT(ASCP)
IHC/ISH Clinical & Research Laboratory
UWHC
600 Highland Ave. DB1-223, Mail Code 3224
Madison, WI 53792
(608)265-6596
Fax:(608)262-7174
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Sharon.Davis-Devine
Sent: Wednesday, February 03, 2010 12:13 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] P63
Do any of you Ventana users have a P63 that is not an RUO? If so, where
do you get it from? Thanks a bunch.
Sharon Davis-Devine, CT (ASCP)
Cytology-Histology Supervisor
Carle Foundation Hospital
Laboratory and Pathology Services
611 West Park Street
Urbana, Illinois 61801
217-383-3572
sharon.davis-devine <@t> carle.com
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------------------------------
Message: 5
Date: Wed, 3 Feb 2010 12:49:28 -0600
From: "Mahoney,Janice A" <Janice.Mahoney <@t> alegent.org>
Subject: [Histonet] RE: GI Biopsies
To: "'Anderson, David W'" <david.w.anderson <@t> Vanderbilt.Edu>,
"histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<8F0EE4144E8E2F4CA1F6B051A2E5BFEE46FADC06 <@t> EXCHMBC2.ad.ah.local>
Content-Type: text/plain; charset="us-ascii"
What kind of stainer do you have?
Jan,
Omaha
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Anderson, David W
Sent: Wednesday, February 03, 2010 12:10 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] GI Biopsies
We are having an issue with our GI Biopsy H&E Stains. On our GI Biopsies, we currently cut eight slides, staining levels 2, 5 and 8 with H&E. The pathologist are noticing that the level two slide looks great but then levels 5 and 8 will have what they call "Blue Blobs" on the tissue sections. They tend to look like formalin salt crystals. Does anyone else experience this? Do any of you have a resolution to this problem? Thank you so much for your help with this matter.
David Anderson,HT(ASCP)
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Message: 6
Date: Wed, 3 Feb 2010 14:01:50 -0500
From: Joyce Cline <Joyce.Cline <@t> wchsys.org>
Subject: RE: [Histonet] Eosin leaching out of sections
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<FE2197D76184F4408CFEBD95C28282685BF0B19B9D <@t> WCHSXCHCM.wchsys.org>
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Humidity has played a large part in our slides leaching Eosin. We have a dehumidifier in our staining room and we also use H2Blue Beads in our Xylene substitute. The beads absorb water from the substitute and help prevent leaching of the stain.
The beads are ordered from Amercian Mastertech.
Joyce Cline, Technical Specialist
Hagerstown Medical Laboratory
301-665-4980
fax 301-665-4941
________________________________________
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Scott
Hendricksen
Sent: 03 February 2010 04:16
To: Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Eosin leaching out of sections
Hi again,
Does anyone have an issue with eosin leaching out of sections a few days
after they have been coverslipped on an H&E stained slide?
Thanks,
Scott Hendricksen HT(ASCP)
_______________________________________________
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Histonet <@t> lists.utsouthwestern.edu
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------------------------------
Message: 7
Date: Wed, 3 Feb 2010 14:11:00 -0500
From: "Rathborne, Toni" <trathborne <@t> somerset-healthcare.com>
Subject: RE: [Histonet] Eosin leaching out of sections
To: "Joyce Cline" <Joyce.Cline <@t> wchsys.org>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<E78340C766A5284D999F5F5891DDF8900BAF91D8 <@t> smcmail.somerset-healthcare.com>
Content-Type: text/plain; charset="utf-8"
Where do you get the beads from?
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Joyce
Cline
Sent: Wednesday, February 03, 2010 2:02 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Eosin leaching out of sections
Humidity has played a large part in our slides leaching Eosin. We have a dehumidifier in our staining room and we also use H2Blue Beads in our Xylene substitute. The beads absorb water from the substitute and help prevent leaching of the stain.
The beads are ordered from Amercian Mastertech.
Joyce Cline, Technical Specialist
Hagerstown Medical Laboratory
301-665-4980
fax 301-665-4941
________________________________________
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Scott
Hendricksen
Sent: 03 February 2010 04:16
To: Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Eosin leaching out of sections
Hi again,
Does anyone have an issue with eosin leaching out of sections a few days
after they have been coverslipped on an H&E stained slide?
Thanks,
Scott Hendricksen HT(ASCP)
_______________________________________________
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Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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Message: 8
Date: Wed, 3 Feb 2010 13:28:07 -0600
From: "Sharon.Davis-Devine" <Sharon.Davis-Devine <@t> carle.com>
Subject: [Histonet] P63 RUO
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<50003EC02E2CEA4583BEB3CD08EAC1E090DDF1 <@t> EXCHANGEBE2.carle.com>
Content-Type: text/plain; charset="us-ascii"
Another question for the group of experts. Is it true that you cannot
bill for antibodies that are classified as a RUO for Ventana?
Sharon Davis-Devine, CT (ASCP)
Cytology-Histology Supervisor
Carle Foundation Hospital
Laboratory and Pathology Services
611 West Park Street
Urbana, Illinois 61801
217-383-3572
sharon.davis-devine <@t> carle.com
------------------------------
Message: 9
Date: Wed, 3 Feb 2010 15:03:50 -0500
From: "McMahon, Loralee A" <Loralee_Mcmahon <@t> URMC.Rochester.edu>
Subject: [Histonet] RE: P63 RUO
To: "Sharon.Davis-Devine" <Sharon.Davis-Devine <@t> carle.com>,
"histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<C27AA2A01CEF31469813089E226F582E3D7CE57C <@t> urmcms7.urmc-sh.rochester.edu>
Content-Type: text/plain; charset="us-ascii"
It doesn't matter what machine you run them on or if you do them by hand. You cannot bill for anything that is RUO. ASR and IVD's muct be properly validated before you bill for them as well.
Loralee McMahon, HTL (ASCP)
Immunohistochemistry Supervisor
Strong Memorial Hospital
Department of Surgical Pathology
(585) 275-7210
________________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sharon.Davis-Devine [Sharon.Davis-Devine <@t> carle.com]
Sent: Wednesday, February 03, 2010 2:28 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] P63 RUO
Another question for the group of experts. Is it true that you cannot
bill for antibodies that are classified as a RUO for Ventana?
Sharon Davis-Devine, CT (ASCP)
Cytology-Histology Supervisor
Carle Foundation Hospital
Laboratory and Pathology Services
611 West Park Street
Urbana, Illinois 61801
217-383-3572
sharon.davis-devine <@t> carle.com
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------------------------------
Message: 10
Date: Wed, 03 Feb 2010 16:20:03 -0500
From: mtitford <@t> aol.com
Subject: [Histonet] Microarray info thanks!
To: Histonet <@t> pathology.swmed.edu
Message-ID: <8CC732DC90E018F-1D5C-2A59 <@t> webmail-m095.sysops.aol.com>
Content-Type: text/plain; charset="us-ascii"
Thank you everyone for all the information about microarray equipment. As usual on the Histonet, everyone was quick with the information, generous with their time, and informative.
Michael Titford
USA Pathology
Mobile AL USA
------------------------------
Message: 11
Date: Wed, 3 Feb 2010 14:04:20 -0800 (PST)
From: Jeffrey Silverman <pathmaster <@t> yahoo.com>
Subject: [Histonet] Drying ovens, eosin fading, microtome PM, eyeballs
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID: <333638.6266.qm <@t> web111109.mail.gq1.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Our routine sections, picked up on regular slides with no adhesive in the water bath, dry at 65C for 15 minutes in the oven on our Leica stainer XL, we never lose sections. Sections for IHC and specials go on Plus slides and dry in an oven also at 65C for at least 1 hour. Then they get the same Leica XL oven drying cycle before deceration.
Fading eosin usually means incomplete dehydration, trace water in the xylene etc.
Belair does all our PM's, microtomes, cryostats, they've kept our 30 year old VIP running despite the fact that Sakura no longer supports the oldest models (like ours).
Eyeballs should not be opened until they have been fixed for at least two days intact in 10% buffered formalin. Then, using a brand new blade, slice off the two callottes, proceeding from cornea posteriorly to the optic nerve, from a central section which includes the optic nerve and the pupil. Some retinal separation is inevitable, this technique will minimize it.
Jeffrey Silverman, HT HTL QIHC
Pathologists' Assistant
Southside Hospital NSLIJHS
Bay Shore, New York USA
------------------------------
Message: 12
Date: Wed, 3 Feb 2010 17:10:38 -0500
From: "Sherwood, Margaret " <MSHERWOOD <@t> PARTNERS.ORG>
Subject: Re: [Histonet] TRAP assay - acid phosphatase
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<073AE2BEA1C2BA4A8837AB6C4B943D9703E23FDC <@t> PHSXMB30.partners.org>
Content-Type: text/plain; charset="us-ascii"
Has anybody used the TRAP assay kit from Sigma for bone? We want to use it on
mouse tibia for IHC.
If so, 1) can you do it on paraffin and/or frozen sections?
2) if you do it on frozens, do you decalcify first?
3) what type of blade do you use for sectioning on the cryostat?
3) what procedure do you follow after sectioning: do you fix slides
(i.e. acetone, etc.)
Thank you.
Peggy
Peggy Sherwood
Lab Associate, Photopathology
Wellman Center for Photomedicine (EDW 214)
Massachusetts General Hospital
55 Fruit Street
Boston, Massachusetts 02114-2696
617-724-4839 (voice mail)
617-726-6983 (lab)
617-726-1206 (fax)
msherwood <@t> partners.org
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Message: 13
Date: Wed, 3 Feb 2010 14:47:47 -0800 (PST)
From: kyle ayres <ayreskyle <@t> yahoo.com>
Subject: [Histonet] slide drying
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID: <47296.74380.qm <@t> web31907.mail.mud.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Place the slides in every other slot of basket and dry for 15-20 min at 69 celcius, let cool in front of fan for 1-2 min. You are able to combine baskets at this step.
Kyle HT BS
Nacogdoches Memorial
Our lab just received a slide drying oven, and we are trying to figure
out what's a good temperature for the slides to be heated and for how
long. We are mainly doing H&E slides.
The way we are currently drying them is under a small fan, then heat
them up, and back to the fan. Then finally heating them up and off to
the stainer.
Just putting them in the slide drying oven for 10 min at 80C melts the
paraffin, but leaves some water on the bottom of some sections.
Any suggestions to get the slides dry asap ? Also is the oven much help
to your labs?
Thanks !
Scott Hendricksen HT(ASCP)
------------------------------
Message: 14
Date: Wed, 3 Feb 2010 15:48:31 -0700
From: "Liz Chlipala" <liz <@t> premierlab.com>
Subject: RE: [Histonet] TRAP assay - acid phosphatase
To: "Sherwood, Margaret " <MSHERWOOD <@t> PARTNERS.ORG>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<EE33BE5C905A3046A7FF8F58A64C8E4B0E74FF <@t> server.PremierLab.local>
Content-Type: text/plain; charset="iso-8859-1"
We have used it on FFPE EDTA decaled mouse bone, with a little bit of fiddling with the protocol it works pretty good
Liz
________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Sherwood, Margaret
Sent: Wed 2/3/2010 3:10 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] TRAP assay - acid phosphatase
Has anybody used the TRAP assay kit from Sigma for bone? We want to use it on
mouse tibia for IHC.
If so, 1) can you do it on paraffin and/or frozen sections?
2) if you do it on frozens, do you decalcify first?
3) what type of blade do you use for sectioning on the cryostat?
3) what procedure do you follow after sectioning: do you fix slides
(i.e. acetone, etc.)
Thank you.
Peggy
Peggy Sherwood
Lab Associate, Photopathology
Wellman Center for Photomedicine (EDW 214)
Massachusetts General Hospital
55 Fruit Street
Boston, Massachusetts 02114-2696
617-724-4839 (voice mail)
617-726-6983 (lab)
617-726-1206 (fax)
msherwood <@t> partners.org
The information in this e-mail is intended only for the person to whom it is
addressed. If you believe this e-mail was sent to you in error and the e-mail
contains patient information, please contact the Partners Compliance HelpLine at
http://www.partners.org/complianceline . If the e-mail was sent to you in error
but does not contain patient information, please contact the sender and properly
dispose of the e-mail.
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------------------------------
Message: 15
Date: Wed, 3 Feb 2010 17:13:43 -0600
From: "Delaney, Sondra L" <Sondra.Delaney <@t> Integris-Health.com>
Subject: [Histonet] FW: FFPE Microwave processed biopsies
To: "'histonet <@t> lists.utsouthwestern.edu'"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<70F29E1F984F6D45A07F071AFAAFC6D308C1AFCA63 <@t> INTEGRISMAIL.corp.integris-health.com>
Content-Type: text/plain; charset="us-ascii"
-----Original Message-----
From: Chad Miller [mailto:cmiller <@t> labmd.org]
Sent: Wednesday, February 03, 2010 4:46 PM
To: Delaney, Sondra L
Cc: Chad Miller
Subject: FFPE Microwave processed biopsies
Microtomy Issue:
Our lab has recently switched from using GTF (Glycol Tissue Fixative) as a primary fixative to10%NBF for prostate biopsies. I am experiencing issues with wrinkles, to varying degrees, along the length of the biopsies especially around the lumens of glandular structures. Sections are taken at 3 micrometers with zero visible compression/wrinkling/washboarding etc... of the tissue. When the ribbon is placed on the water bath (47-50 C) the tissue seems to contract and wrinkling appears although, only in the tissue....similar to the coils of a compressed slinky. I have adjusted the water bath temp. increasingly higher until my wax can no longer stand the temp and dissipates around the tissue. This has not cured the wrinkling problem but has shown some improvement. Prostate biopsies that have been fixed in GTF and processed identically to the formalin fixed biopsies are simply perfect in every way. Specimens are embedded using Polyfin (TBS, M.P. 55C) and kept cool for sectioning on
ice trays. Sections are collected on charged Histobond slides. Wrinkles appear whether processing solutions are brand new or have been used for previous runs. If anyone has any suggestions ideas or experiences I would sincerely appreciate them. I have included processing times below.
Specimen Type: Prostate Needle Core Biopsy
Processor: Milestone Histos 5 (Microwave) Specimen Fixative: 10%NBF Specimen Prep: Aligned in linear fashion and sandwiched between sponges to maintain orientation.
Max Block Volume: 110 blocks/run
Offline Prep:
1. 70% ETOH Rinse (Manual @ 5minutes)--(Changed every 450 blks)
2. 95% ETOH Rinse (Manual @ 5minutes)--(Changed every 450 blks)
3. 100% ETOH Rinse (Manual @ 5minutes)--(Changed every 450 blks)
Histos 5 Processor: (0-55blk processing protocol)
1. 100% ETOH (15 minutes @ 65 C)--(Changed every 220 blks)
2. Isopropyl (16 minutes @ 68 C)--(Changed every 220 blks)
3. Vacuum Step (2 minutes ambient)
4. Paraffin (Paraplast M.P.56 C) (23 minutes @ 68 C with Vacuum)-(Changed every 450 blks)
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------------------------------
Message: 16
Date: Wed, 3 Feb 2010 16:11:13 -0800
From: Nicole Collette <collette2 <@t> mail.llnl.gov>
Subject: [Histonet] autofluorescence experiment
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID: <p06230909c78fb7baa4ac@[128.115.150.224]>
Content-Type: text/plain; charset="us-ascii" ; format="flowed"
Hello, All,
I am working on doing some IF stains with bone samples (lucky me!). I
am having a difficult time sometimes to assess the antibody since the
autofluorescence gets in the way. I am using undecalcified, FFPE
sections (late embryo and neonate mouse bones). Without treatment, I
see autofluorescence everywhere, but most frustrating is the red
blood cells and the mineralized matrix. It took me a while to get
samples that are properly fixed, section well, and stay on the
slides, so I am not particularly jazzed about messing with the
fixation protocol. Thus, I conducted an experiment today of several
published and voodoo methods to reduce autofluorescence with samples
that did NOT undergo the IF protocol :
no treatment
photobleaching, fluorescent light box, up to 2 weeks
photobleaching, UV crosslinking light, 2 inches from source, 24h
10mM copper sulfate in 50mM ammonium acetate, pH 5.0
0.25% (v/v) NH3 in 70% ethanol (in water)
0.25% (v/v) NH4OH in 70% ethanol (in water) (since it was unclear
from the published reference what the source of ammonia was, and I
have made this mistake before on some other thing)
0.3% (w/v) Sudan Black in 70% ethanol (in water)
I found that the most effective treatment in my hands is Sudan Black
for cell-based autofluorescence, but it did not seem to impact the
autofluorescence from the mineralized matrix at all, while copper
sulfate had significant impact on the autofluorescence in mineralized
bone, but did not quench cell-based autofluorescence as well as the
Sudan Black (it had an even but incomplete impact over the entire
section). I have tried Sudan Black on my slides before, and have
found that it did not seem to interfere significantly with the
antibody signal. I modified the Sudan Black protocol to eliminate the
"goopies" and "chunkies" resulting on my slides from previous
attempts, and am happy with the results- a light, even stain.
My question to all you chemistry folks: Is there some reason why
copper sulfate treatment followed by washing and subsequent Sudan
Black treatment (and more washing) cannot or should not be used? I
tried them together on my unstained slides, they look pretty darn
fabulous. Just striving for clean data and beautiful pictures.
Thanks again for all your help,
Sincerely,
Nicole Collette
Lawrence Livermore National Lab/ UC Berkeley
------------------------------
Message: 17
Date: Wed, 3 Feb 2010 17:42:18 -0800 (PST)
From: soofia siddiqui <soofias2 <@t> yahoo.com>
Subject: Re: [Histonet] Derm Lab
To: histonet <@t> lists.utsouthwestern.edu, Eric Sulkosky
<esulkosky <@t> gmail.com>
Message-ID: <692051.1111.qm <@t> web112503.mail.gq1.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Raj,
If you answer Eric's email I may help you too, because I set up a highly complex testing lab at dermatology starting from scratch with an empty
room when they were starting the remodeling the room. The room may have already been equipped with ventilation, plumbing and
electrical but they still had to put outlets and design the working are and bench surface.
Soofia
--- On Wed, 12/30/09, Eric Sulkosky <esulkosky <@t> gmail.com> wrote:
From: Eric Sulkosky <esulkosky <@t> gmail.com>
Subject: [Histonet] Derm Lab
To: histonet <@t> lists.utsouthwestern.edu
Date: Wednesday, December 30, 2009, 8:40 AM
RAJ,
What tips are you looking for? Are you starting from scratch with an empty
room or has the room already been equipped with ventilation, plumbing and
electrical?
Eric
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 18
Date: Wed, 3 Feb 2010 22:11:03 -0800 (PST)
From: soofia siddiqui <soofias2 <@t> yahoo.com>
Subject: [Histonet] Frozen Sections Slides per Day Question
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID: <552462.75622.qm <@t> web112516.mail.gq1.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Hi dear histology experts,
I will greatly appreciate if somebody can let me know the estimated number of slides (with 3 sections per slide) per day on average a lab technician can cut (of frozen tissue).
I am a lab technician ( not a histotech) and work alone in a highly complex testing specialzed low volume dermatology lab.My job description includes 30 % of immunohistochemistry related dutites. I cut skin frozen sections and do immunohistochemistry manually for several years . My routiene panel consists of 12 antibodies for T-cells surface markers. Ocassionally I add another panel of 8 antibodies for B-Cells.
I am very slow in cutting sections and strugle a lot to get good sections.
If I spend entire day ( 8 hours) just cutting 3-4 section on each slide with slow speed. What number of slides should be considered as efficent cutting?
What number of blocks (12 slides for each block with 3-4 sections) should I finish in one day?
Help me please if you can. Thanks. Soofia
------------------------------
Message: 19
Date: Thu, 4 Feb 2010 05:48:38 -0500
From: "Mary Lloyd" <lloyd.3 <@t> osu.edu>
Subject: [Histonet] mailers
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<04A71EFE05FE5F4DAA05D0500FD0597002A014F1 <@t> Distal.dentnet.dent.ohio-state.edu>
Content-Type: text/plain; charset="us-ascii"
We have been using a round double mailer with a metal insert from Fisher
Scientific. The size for the outside cardboard mailer is 2 3/8" by 5
3/4 inch. Fisher has discontinued this mailer. Does anyone know a
reasonable company that would carry these mailers. I found a company
that has a similar product that is larger, but our cost for mailing will
increase to mail the containers. I would appreciate any help. Thanks
------------------------------
Message: 20
Date: Thu, 04 Feb 2010 09:15:19 -0500
From: "Percival Karen" <KPercival <@t> wyeth.com>
Subject: RE: [Histonet] Tape/Film Coverslips vs. Glass
To: <histonet <@t> lists.utsouthwestern.edu>, <Cathy.Crumpton <@t> tuality.org>,
"Pyse Cynthia" <cpyse <@t> x-celllab.com>
Message-ID: <4B6A902702000011001F84DC <@t> gv01a67m.gv.us.pri.wyeth.com>
Content-Type: text/plain; charset=US-ASCII
I've used both over the years, and I love the tape coverslippers. The slides dry very quickly because there is no mounting media step. The tape sticks to the slide via a chemical reaction between the xylene and the tape. One thing to remember with tape, you cannot use recycled xylene as your last xylene step before coverslipping. If I remember correctly, you cannot use xylene substitutes either. It's been a while, so that may have changed.
The tape coverslips are easily removed with acetone.
The pathologists loved it because they received their slides immediately. We didn't have to allow drying time before handing them in.
I didn't find any tissue fading over time, either.
Just my opinion. Good luck.
>>> "Cynthia Pyse" <cpyse <@t> x-celllab.com> 2/3/2010 9:21 AM >>>
Cathy
I prefer the glass coverslips. It seems to give more protection to the
tissue. Recoverslipping is also easier than with the tape. Just my opinion.
Hope this helps.
Cindy Pyse, CLT, HT (ASCP)
Histology Supervisor
X-Cell Laboratories
e-mail cpyse <@t> x-celllab.com
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Cathy.Crumpton <@t> tuality.org
Sent: Monday, February 01, 2010 11:40 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Tape/Film Coverslips vs. Glass
Hello all,
We mig=t be getting a new coverslipper and the pathologists asked me
to get feedb=ck from my fellow histotechs that have used both glass
and tape coverslips=. Which type is most prefered for general
histology and cytology?&nbs=p; I did some research in the archives
and most of the responses were dated=004. Has anything changed
about the tape systems since then? =I would be interested in
receiving your input.
Thanks,
Cath=
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------------------------------
Message: 21
Date: Thu, 4 Feb 2010 10:05:30 -0500
From: rgrow <@t> bmnet.com
Subject: [Histonet] Re: Eosin leaching out of sections
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<OF31DDCF3C.906DA047-ON852576C0.0052675F-852576C0.0052E72F <@t> bmnet.com>
Content-Type: text/plain; charset=US-ASCII
Hi Scott,
Your discription of eosin leaching out after a few days is classic for the
possibility of bleach contamination on your staining containers. There are
multiple possibilities of other causes, but this could be a simple cause,
easily solved.
Renee Grow, BA., HT (ASCP)
rgrow <@t> bmnet.com
Histology Supervisor
Blount Memorial Hospital
907 E. Lamar Alexander Pkwy.
Maryville, TN 37804-5016
(865) 977-4744
(865) 977-5766 Fax
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Scott
Hendricksen
Sent: 03 February 2010 04:16
To: Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Eosin leaching out of sections
Hi again,
Does anyone have an issue with eosin leaching out of sections a few days
after they have been coverslipped on an H&E stained slide?
Thanks,
Scott Hendricksen HT(ASCP)
_______________________________________________
------------------------------
Message: 22
Date: Thu, 4 Feb 2010 10:19:43 -0500
From: "Sherwood, Margaret " <MSHERWOOD <@t> PARTNERS.ORG>
Subject: Re: [Histonet] TRAP Assay and IHC
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<073AE2BEA1C2BA4A8837AB6C4B943D9703E23FDE <@t> PHSXMB30.partners.org>
Content-Type: text/plain; charset="us-ascii"
I want to thank everyone who responded to my inquiry re: TRAP Assay. I don't
think I made my request that clear re: IHC. We want to do both TRAP Assay and
IHC (separately, of course) on mouse tibia.
We would like to use frozen sections of mouse tibia for IHC. Our initial try to
cut tibia on the cryostat has resulted in less than optimal sections. It
appears that the tissue is lifting out of the section.
In re: IHC on frozens - 1) do you treat the tibia before embedding in OCT?
2) what knife do you cut with on the cryostat? 3) do you fix slides with cold
acetone before the IHC procedure (we ususally do)?
Thanks again!
Peggy
Peggy Sherwood
Lab Associate, Photopathology
Wellman Center for Photomedicine (EDW 214)
Massachusetts General Hospital
55 Fruit Street
Boston, Massachusetts 02114-2696
617-724-4839 (voice mail)
617-726-6983 (lab)
617-726-1206 (fax)
msherwood <@t> partners.org
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------------------------------
Message: 23
Date: Thu, 4 Feb 2010 09:18:54 -0600
From: "Rosa Fields" <rfields <@t> gidocs.net>
Subject: RE: [Histonet] Tape/Film Coverslips vs. Glass
To: "Percival Karen" <KPercival <@t> wyeth.com>,
<histonet <@t> lists.utsouthwestern.edu>, <Cathy.Crumpton <@t> tuality.org>,
"Pyse Cynthia" <cpyse <@t> x-celllab.com>
Message-ID:
<07732CE52EC3174AB891DE1C62DB4D8FC7B6A3 <@t> GIEXCHANGE.gidocs.net>
Content-Type: text/plain; charset="iso-8859-1"
The tape cover slippers are quicker, easier to manage the slides, but the cover slips pop off over time; the glass cover slippers are a bit temperamental; the slides take longer to dry, and the quality is better, and the cover slip stays on.
Rosa Fields, HT (ASCP)
Gastroenterology Specialties
Histology Supervisor
4545 R Street
Lincoln, NE 68503
402-465-4545
rfields <@t> gidocs.net
The information contained in the message and the documents accompanying this message contain information that is privileged and confidential and is intended only for the use of the individual or entity named above. If the reader of this message is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication, other than its return to the sender, is strictly prohibited.
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Percival Karen
Sent: Thursday, February 04, 2010 8:15 AM
To: histonet <@t> lists.utsouthwestern.edu; Cathy.Crumpton <@t> tuality.org; Pyse Cynthia
Subject: RE: [Histonet] Tape/Film Coverslips vs. Glass
I've used both over the years, and I love the tape coverslippers. The slides dry very quickly because there is no mounting media step. The tape sticks to the slide via a chemical reaction between the xylene and the tape. One thing to remember with tape, you cannot use recycled xylene as your last xylene step before coverslipping. If I remember correctly, you cannot use xylene substitutes either. It's been a while, so that may have changed.
The tape coverslips are easily removed with acetone.
The pathologists loved it because they received their slides immediately. We didn't have to allow drying time before handing them in.
I didn't find any tissue fading over time, either.
Just my opinion. Good luck.
>>> "Cynthia Pyse" <cpyse <@t> x-celllab.com> 2/3/2010 9:21 AM >>>
Cathy
I prefer the glass coverslips. It seems to give more protection to the
tissue. Recoverslipping is also easier than with the tape. Just my opinion.
Hope this helps.
Cindy Pyse, CLT, HT (ASCP)
Histology Supervisor
X-Cell Laboratories
e-mail cpyse <@t> x-celllab.com
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Cathy.Crumpton <@t> tuality.org
Sent: Monday, February 01, 2010 11:40 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Tape/Film Coverslips vs. Glass
Hello all,
We mig=t be getting a new coverslipper and the pathologists asked me
to get feedb=ck from my fellow histotechs that have used both glass
and tape coverslips=. Which type is most prefered for general
histology and cytology?&nbs=p; I did some research in the archives
and most of the responses were dated=004. Has anything changed
about the tape systems since then? =I would be interested in
receiving your input.
Thanks,
Cath=
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Histonet mailing list
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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------------------------------
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