RE: [Histonet] RE:H&E on GMA sections
Leslie,
You are going to have a hard time getting 1-2 micron GMA sections to look
just like 4-6 micron ffpe sections with H&E. Besides the obvious thickness
difference the stain has to pass thru the GMA as it is not removed. In my
experience this takes longer with more concentrated stain. I use Gill's#3
hematoxylin for 15min. wash in tap h20 to achieve bluing. I use a
commercial alcoholic eosin/phloxine. After Hematox. And wash I airdry the
sections standing up (10-20 min. in my dry climate) I then dip briefly in
95% alcohol, dip about 90 times (I keep it moving) in the Eosin, then
quickly thru 2 chgs. Of 95 ETOH, about 5 dips ea., then 2 chgs. Of 100%etoh,
once you get past the alcohol with water in it you can slow down, it is the
water/alcohol mix that causes GMA to lift off the slide in my experience, I
usually go into 100 about 1 min. ea chg then in 3 xylenes and permount.
This is the only stain I do for GMA that I don't airdry the sections and
coverslip with just a brief dip in xylene, the reason I use alcohol for the
HE is because I use an alcoholic eosin, even when I use aqueous eosin I go
thru Etoh (albeit quickly) to get the differentiation of the eosin we
desire. Every other stain I do for GMA, if it ends in water, I airdry the
section, dip once in xylene and permount.
Patsy
Patsy Ruegg, HT(ASCP)QIHC
IHCtech
12635 Montview Blvd. #216
Aurora, CO 80010
720-859-4060
fax 720-859-4110
pruegg@ihctech.net
www.ihctech.net
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Carl Hobbs
Sent: Saturday, March 25, 2006 11:47 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE:H&E on GMA sections
Well.....after leaving my slide-mounted GMA-embedded sections in a 60C oven
for an hour. I'll rinse them in tapwater and then leave them in Hx for about
20mins. After that, I'll rinse them in tapwater, have a look down the
m'scope to note the intensity of Hx staining and differentiate in 0.5% HCL
in 70% Alc ( water if you want)until I think that the balance is good. Then
I will leave in 0.5% Eosin for as long as I want ( say, 20mins). Then wash
in tapwater until I achieve the best Hx-Eosin balance. I then blot dry my
resin sections and place them in the same oven to dry...min an hour. Then
put them in xylene and , after 15-odd mins ..mount in DPX!
No times are critical, IMHO. That's the great thing about H&E....just get a
routine that is reproducable/consistent for your lab.
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of
histonet-request@lists.utsouthwestern.edu
Sent: None
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 28, Issue 41
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Today's Topics:
1. Staining Glycol Methacrylate (Leslie D Duncan)
2. Ventana IVIEW Detection Kits (Tracey.Lenek@CLS.ab.ca)
3. endothelial marker FFPE Rat (Steven Coakley)
4. RE: anti-human TNFR-1 anti-mouse TNFR-1 and 2 (Guillermo Palao)
5. RE: endothelial marker FFPE Rat (Elizabeth Chlipala)
6. RE: Ventana IVIEW Detection Kits (Sebree Linda A.)
7. peripheral axons in soft tissue (Elizabeth Chlipala)
8. RE: endothelial marker FFPE Rat (Trajkovic, Dusko)
9. Job openings in Michigan (Mary McCoy)
10. RE: peripheral axons in soft tissue (Smith, Allen)
11. RE: Input on Milestone RHS-1 and RHS-2 (Justin Thomas)
12. Autodial alarms (Marian powers)
13. Hybrid capture, CISH (Katia Cristina Catunda)
14. Xylene exposure (Jeannie Heck)
15. Re: Xylene exposure (Rene J Buesa)
----------------------------------------------------------------------
Message: 1
Date: Fri, 24 Mar 2006 12:28:22 -0600
From: Leslie D Duncan
Subject: [Histonet] Staining Glycol Methacrylate
To: histonet@lists.utsouthwestern.edu
Message-ID: <44243A46.3050508@bms.com>
Content-Type: text/plain; charset="iso-8859-1"
Hi,
I am experimenting with glycol methacrylate embedded rodent testes. I
am looking for a good H&E staining protocol to use that will give
results comparable to paraffin-embedded tissue (per request of my
management). We have dehydrated on the tissue processor to 100%
ethanol, ilfiltrated and embedded with JB-4, and sectioned at 1-2
microns. The sections are placed on Poly L-lysine slides. Any
protocols, ideas, and/or suggestions will be greatly appreciated!
Thank you,
Leslie Duncan
Bristol-Myers Squibb, PRI
------------------------------
Message: 2
Date: Fri, 24 Mar 2006 11:35:08 -0700
From:
Subject: [Histonet] Ventana IVIEW Detection Kits
To:
Message-ID:
<4BC300747AF87A48BCDF8E48BC2885CE02C1D598@mail1.calgary.com>
Content-Type: text/plain; charset="iso-8859-1"
Just a general question for all the Ventana automated IHC equipment users -
this time last year we found that the IVIEW detection kits were
overstaining. Now we are experiencing weak staining. Has anyone else found
variabilities with the staining in their kits? We have relayed our issues
to Ventana but as usual are the only ones complaining.
Tracey
CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain
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------------------------------
Message: 3
Date: Fri, 24 Mar 2006 10:08:33 -0800 (PST)
From: Steven Coakley
Subject: [Histonet] endothelial marker FFPE Rat
To: Histonet@lists.utsouthwestern.edu
Message-ID: <20060324180833.66058.qmail@web38202.mail.mud.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Has anyone any knowledge of any endothelial markers, such as CD31, RECA,
ect....
for FFPE for a rat muscle angiogenesis study.
Steve
---------------------------------
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------------------------------
Message: 4
Date: Fri, 24 Mar 2006 20:12:57 +0100 (CET)
From: Guillermo Palao
Subject: [Histonet] RE: anti-human TNFR-1 anti-mouse TNFR-1 and 2
To: Histonet
Message-ID: <20060324191258.74730.qmail@web26210.mail.ukl.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Again, any suggestions for good commercially available antibody for mouse
TNRs 1 and 2 (CD120 a,b) are most welcome.
Thanks for all previous and future help in saving time with painful
optimizations of non-working antibodies.
Guillermo
Alessandro Serra escribis:
Dear All
Has anybody used succesfully any commercially available anti-human
TNFR-1 (CD120a)?
If yes, could you please be so kind to tell me which one?
Thank you very much for your help
Alessandro Serra, BSc.
---------------------------------
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------------------------------
Message: 5
Date: Fri, 24 Mar 2006 12:31:52 -0700
From: "Elizabeth Chlipala"
Subject: RE: [Histonet] endothelial marker FFPE Rat
To: "'Steven Coakley'" ,
Message-ID: <000a01c64f79$9a72f4e0$0300a8c0@Chlipala>
Content-Type: text/plain; charset="iso-8859-1"
Steve
I have used the goat-anti mouse PECAM-1 from santa cruz for rat tissue
and it works. The only thing is that I happen to have the older lot
number that works in FFPE tissues. I don't know what newer lots will
do.
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
P.O. Box 18592
Boulder, Colorado 80308
Office: (303) 735-5001
Fax: (303) 735-3540
liz@premierlab.com
www.premierlab.com
Ship to Address:
Premier Laboratory
University of Colorado
MCDB, Room A3B40
Boulder, Colorado 80309
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steven
Coakley
Sent: Friday, March 24, 2006 11:09 AM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] endothelial marker FFPE Rat
Has anyone any knowledge of any endothelial markers, such as CD31, RECA,
ect....
for FFPE for a rat muscle angiogenesis study.
Steve
---------------------------------
Blab-away for as little as 1"/min. Make PC-to-Phone Calls using Yahoo!
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------------------------------
Message: 6
Date: Fri, 24 Mar 2006 13:36:22 -0600
From: "Sebree Linda A."
Subject: RE: [Histonet] Ventana IVIEW Detection Kits
To: ,
Message-ID:
Content-Type: text/plain; charset="US-ASCII"
We have not experienced that phenomenon ever that I can recall.
Linda Sebree, HT(ASCP)
University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
A4/204-3224
600 Highland Ave.
Madison, WI 53792
(608)265-6596
FAX: (608)262-7174
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
Tracey.Lenek@CLS.ab.ca
Sent: Friday, March 24, 2006 12:35 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Ventana IVIEW Detection Kits
Just a general question for all the Ventana automated IHC equipment
users - this time last year we found that the IVIEW detection kits were
overstaining. Now we are experiencing weak staining. Has anyone else
found variabilities with the staining in their kits? We have relayed
our issues to Ventana but as usual are the only ones complaining.
Tracey
CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may
contain confidential, personal and/or privileged information intended
for a specific purpose and recipient. If you are not the intended
recipient do not disclose, copy, retain, distribute, use or modify any
of the contents of this transmission. If you received this transmission
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Thank you.
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------------------------------
Message: 7
Date: Fri, 24 Mar 2006 12:40:43 -0700
From: "Elizabeth Chlipala"
Subject: [Histonet] peripheral axons in soft tissue
To:
Message-ID: <000001c64f7a$d6a91f10$0300a8c0@Chlipala>
Content-Type: text/plain; charset="us-ascii"
Hello Everyone
Is there a stain that I can use that will detect peripheral nerve
(axons) in soft tissue. I have checked around a bit and it looks like
performing a Bodian would work. I would appreciate any input or advice
on this.
Thanks in advance
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
P.O. Box 18592
Boulder, Colorado 80308
Office: (303) 735-5001
Fax: (303) 735-3540
liz@premierlab.com
www.premierlab.com
Ship to Address:
Premier Laboratory
University of Colorado
MCDB, Room A3B40
Boulder, Colorado 80309
------------------------------
Message: 8
Date: Fri, 24 Mar 2006 11:46:35 -0800
From: "Trajkovic, Dusko"
Subject: RE: [Histonet] endothelial marker FFPE Rat
To: "Steven Coakley" ,
Message-ID:
<3AD0BD3142459B4E9B12CBEAFF2B89B202A94F4F@lajamrexm01.amer.pfizer.com>
Content-Type: text/plain; charset="iso-8859-1"
I had Santa Cruz goat antibody PECAM-1 sc-1505 (lot# G1103) that worked well
on rat tissue, however the new lot number (C2405) which I received last
week, is not working well at all. I will be getting a new lot number from
them next Tuesday. I will post my results later in the week.
Dusko Trajkovic
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Elizabeth
Chlipala
Sent: Friday, March 24, 2006 11:32 AM
To: 'Steven Coakley'; Histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] endothelial marker FFPE Rat
Steve
I have used the goat-anti mouse PECAM-1 from santa cruz for rat tissue
and it works. The only thing is that I happen to have the older lot
number that works in FFPE tissues. I don't know what newer lots will
do.
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
P.O. Box 18592
Boulder, Colorado 80308
Office: (303) 735-5001
Fax: (303) 735-3540
liz@premierlab.com
www.premierlab.com
Ship to Address:
Premier Laboratory
University of Colorado
MCDB, Room A3B40
Boulder, Colorado 80309
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steven
Coakley
Sent: Friday, March 24, 2006 11:09 AM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] endothelial marker FFPE Rat
Has anyone any knowledge of any endothelial markers, such as CD31, RECA,
ect....
for FFPE for a rat muscle angiogenesis study.
Steve
---------------------------------
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----------------------------------------------------------------------
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------------------------------
Message: 9
Date: Fri, 24 Mar 2006 15:18:53 -0500
From: "Mary McCoy"
Subject: [Histonet] Job openings in Michigan
To: histonet@lists.utsouthwestern.edu
Cc: Marianna Cerecke
Message-ID:
Content-Type: text/plain; charset=us-ascii
Lakeland Regional Health System
Position: Histotechnologist
Salary: Competitive
Place of work: Lakeland Hospital, St. Joseph, MI
Position status: Two, full-time permanent positions
Number of Vacant Positions: 02
Description of position:
Performs technical functions necessary in preparation of tissues for
microscopic examination by the pathologist; assists the coordinator with
quality assurance monitoring; management of the immunohistochemical
(IHC) stains and associated quality control; assists the coordinator
with the implementation of new procedures and the evaluation of new
instruments; and assists the coordinator with training as necessary. HT
or HTL (ASCP) certification is preferred; we will consider a Bachelors
degree in a related field and training in the department.
Lakeland offers a comprehensive benefits package, competitive salaries,
PTO, educational reimbursement, and much more. Lakeland is located on
the shores of beautiful Lake Michigan and is just 90 minutes from
Chicago.
Please send cover letter and resume to Marianna Cerecke, Lakeland
Regional Health System, 6418 Deans Hill Rd, Berrien Center, MI 49102.
Or fax to: 269-473-3069. For questions call 269-473-3069 or visit our
website at www.lakelandhealth.org to apply on-line.
------------------------------
Message: 10
Date: Fri, 24 Mar 2006 16:08:38 -0500
From: "Smith, Allen"
Subject: RE: [Histonet] peripheral axons in soft tissue
To: "Elizabeth Chlipala"
Cc: histonet@lists.utsouthwestern.edu
Message-ID:
<5D2189E74151CC42BEC02906BA8996322B91AD@exchsrv01.barrynet.barry.edu>
Content-Type: text/plain; charset="us-ascii"
In my hands, the most reliable technique for demonstrating peripheral nerves
is Kiernan's physical developer method. [pp. 371-372 of the 3rd edition of
J.A. Kiernan's HISTOLOGICAL AND HISTOCHEMICAL METHODS.]
Allen A. Smith, Ph.D.
Professor of Anatomy
Barry University School of Graduate Medical Sciences
Podiatric Medicine and Surgery
Miami Shores, Florida 33161
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Elizabeth
Chlipala
Sent: Friday, March 24, 2006 2:41 PM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] peripheral axons in soft tissue
Hello Everyone
Is there a stain that I can use that will detect peripheral nerve
(axons) in soft tissue. I have checked around a bit and it looks like
performing a Bodian would work. I would appreciate any input or advice
on this.
Thanks in advance
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
P.O. Box 18592
Boulder, Colorado 80308
Office: (303) 735-5001
Fax: (303) 735-3540
liz@premierlab.com
www.premierlab.com
Ship to Address:
Premier Laboratory
University of Colorado
MCDB, Room A3B40
Boulder, Colorado 80309
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------------------------------
Message: 11
Date: Fri, 24 Mar 2006 13:37:00 -0800 (PST)
From: Justin Thomas
Subject: [Histonet] RE: Input on Milestone RHS-1 and RHS-2
To: histonet@lists.utsouthwestern.edu
Message-ID: <20060324213700.4795.qmail@web35704.mail.mud.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
is anyone familiar with the milestone RHS-1 and RHS-2 microwaves? If so,
what are your pros and cons for the two units. Thanks in advance for any
input!
-thomas
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Message: 12
Date: Fri, 24 Mar 2006 17:27:07 -0500
From: "Marian powers"
Subject: [Histonet] Autodial alarms
To:
Message-ID: <000801c64f92$163b1c70$be4dff45@MEGAN>
Content-Type: text/plain; charset="iso-8859-1"
Looking for an autodial alarm for a tissue processor, any recommendations?
Thanks in advance.
Marian
------------------------------
Message: 13
Date: Fri, 24 Mar 2006 21:08:12 -0300
From: "Katia Cristina Catunda"
Subject: [Histonet] Hybrid capture, CISH
To:
Message-ID: <004401c64fa0$35052890$17d0fea9@privatexx>
Content-Type: text/plain; format=flowed; charset="iso-8859-1";
reply-type=original
Dear collegues,
I would like some information about vendors for Hybrid Capture equipments
and supplies as for CISH, can anybody help me (just know Digene and Zymer
respectively).
And how about some tips for Hybridizators? We've found from boekel and zymed
with a great price diferenciation.
Thanks
Katia
------------------------------
Message: 14
Date: Sat, 25 Mar 2006 07:16:17 -0800 (PST)
From: Jeannie Heck
Subject: [Histonet] Xylene exposure
To: Histonet e-mail ?'s
Message-ID: <20060325151617.49980.qmail@web30705.mail.mud.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
What special precautions, if any, does your facility
make for pregnant employees whose job duties involve
the use of, or exposure to xylene?
__________________________________________________
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------------------------------
Message: 15
Date: Sat, 25 Mar 2006 09:29:41 -0800 (PST)
From: Rene J Buesa
Subject: Re: [Histonet] Xylene exposure
To: Jeannie Heck , Histonet e-mail ?'s
Message-ID: <20060325172941.74110.qmail@web61213.mail.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Three in my staff confronted that situation (not simultaneously!!) and I
tried to rotate each to tasks with much less exposure. On the other hand our
monitoring system always came as "0" (13 ppm / hour) which is approximately
9% for xylene STEL and 13% for xylene TWA exposure limits.
I always found that reasignment of tasks is the best solution but it
requires a strongly implemented crosstraining program.
Hope this will help!
Reni J.
Jeannie Heck wrote:
What special precautions, if any, does your facility
make for pregnant employees whose job duties involve
the use of, or exposure to xylene?
__________________________________________________
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End of Histonet Digest, Vol 28, Issue 41
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