[Histonet] Histogel Protocol FROZEN Histonet Digest, Vol 91,
Issue 34
Kathy Bonness
Kathy.Bonness <@t> UTSouthwestern.edu
Mon Jun 27 11:22:25 CDT 2011
Katy (or anyone else who has used Histogel for FROZEN samples),
I am using Histogel mixed with OCT to create a frozen cell pellet. I was successful with doing a single pellet but now I am being asked to do multiple and have found hit some challenges.
I have never done a TMA but that sounds like a great option. We are a research lab but using the Cell Pellet for a normalizing control of IF intensity (for computational analysis normalization for lung tissue) and one of the largest challenges is retaining cell density while still trying to cryoprotect and freeze well with the OCT surrounding it.
Has anyone worked out a method for frozen cell pellets to embedded with multiple cell lines? Most of what I have found has been from FFPE, any suggestions would be appreciated.
Thank you!
Kathy
Kathy M. Bonness, PhD.
251-533-2661
kathy.bonness <@t> utsouthwestern.edu
http://www.linkedin.com/pub/kathy-bonness/6/1a1/931
UTSW Dallas, TX
Green Center for Computational & Systems Biology
Department of Pharmacology
Altschuler/Wu Lab (ND9.214)
________________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] on behalf of histonet-request <@t> lists.utsouthwestern.edu [histonet-request <@t> lists.utsouthwestern.edu]
Sent: Saturday, June 25, 2011 12:00 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 91, Issue 34
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Today's Topics:
1. RE: HistoGel (Milne, Katy)
2. Histogel Problem (Amos Brooks)
3. RE: Histogel Problem (Delossantos_Roseann)
4. bluing (Webb, Dorothy L)
5. Re: bluing (Rene J Buesa)
6. RE: bluing (Harrison, Sandra C.)
7. blades (Webb, Dorothy L)
8. Re: blades (Sean McBride)
9. Re: blades (Esther C Peters)
10. Re: blades (Victoria Baker)
11. Re: blades (Jennifer MacDonald)
12. HT Position - Irvine, CA (Eric Velazquez)
13. Re: HT Position - Irvine, CA (Lee & Peggy Wenk)
14. Contents of Histonet digest (Aurea Marquez)
15. Re: bluing (Lee & Peggy Wenk)
16. Leica Bond for IHCs (Sheila Adey)
17. RE: Bluing (gayle callis)
----------------------------------------------------------------------
Message: 1
Date: Fri, 24 Jun 2011 10:27:09 -0700
From: "Milne, Katy" <kmilne <@t> bccancer.bc.ca>
Subject: [Histonet] RE: HistoGel
To: "'histonet <@t> lists.utsouthwestern.edu'"
<histonet <@t> lists.utsouthwestern.edu>, "'mjdessoye <@t> wvhcs.org'"
<mjdessoye <@t> wvhcs.org>
Message-ID:
<3FEFF18FF4E1914A9AB7D8498591BE8610CF058850 <@t> VEXCCR02.phsabc.ehcnet.ca>
Content-Type: text/plain; charset="us-ascii"
We use histogel a lot in our lab. It's a research lab and we use it for a few purposes - pelleting cultured cells then creating multi-culture TMAs for testing antibodies and also pelleting cells from ascites and pleural effusions. Has also been used to process really small samples that could have been lost in the processor through the cassettes.
Works quite well. The researchers just put the samples in histogel and give it to me in formalin then I process it as I would regular tissue. Cuts very well too.
Katy
Message: 3
Date: Fri, 24 Jun 2011 12:26:46 -0400
From: "Dessoye, Michael J" <mjdessoye <@t> wvhcs.org>
Subject: [Histonet] HistoGel
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<E2547E1CD0EE324488A2940994571EFA0401F3AE <@t> WVHCS-EXCHANGE.wvhcs.com>
Content-Type: text/plain; charset="iso-8859-1"
Hello,
Does anyone out there have any experience with HistoGel? It's Richard Allan/Thermo Fisher. They claim that you can "embed" scant tissues in the gel and then process, embed, and cut as usual. Just wondering how it works in the real world....
Michael J. Dessoye, M.S. | Histology Supervisor | Wyoming Valley Health Care System | mjdessoye <@t> wvhcs.org <mailto:mjdessoye <@t> wvhcs.org> |
575 N. River Street | Wilkes Barre, PA 18764 | Tel: 570-552-1485 | Fax: 570-552-1526
------------------------------
Message: 2
Date: Fri, 24 Jun 2011 13:29:09 -0400
From: Amos Brooks <amosbrooks <@t> gmail.com>
Subject: [Histonet] Histogel Problem
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <BANLkTinbTG=QCS3pEuF8zfa3bz2WqRtheQ <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1
Hi,
I have a problem with some blocks that were prepared with Histogel. I was
hoping someone else might have had a similar problem and figured it out. I
took a photo of the blocks that were mads and put them in a Picassa album
here:
https://picasaweb.google.com/lh/photo/APO3HsIMa2_jPOEs3QRGUjhz3qi22FNPb2i5JJnBCAk?feat=directlink
The long & short of it is that the blocs were prepared by the researcher
for me to process. They are mouse kidneys. Now it is entirely possible for
him to have goofed something up in preparing the Histogel blocks. I wasn't
there when he did it, but when I looked at them before processing, they all
looked fine. (Like the adjacent good one in the photo). When they were
processed they were placed in the VIP right next to each other. When I went
to embed them this morning all but one of the four looked fine. The one that
didn't come out well looked like the Histogel had shrunk up and shriveled
around the kidneys. I am sure this will be aweful to cut, and the researcher
is going to have a bird over it since this happened with another project
previously. I would like to have a decent explanation for him, so if anyone
knows what might have happened and has suggestions I would love to hear it
(yes vendors too are welcome to answer this of course!).
By the way, this was processed on a rather short cycle of 15-20 min per
station of graded ETOH from 70% to 100% with 3 xylene stations and 4
paraffin stations (45 min for these). It seems fine for everything else that
was on the processor. Just that one Histogel block was the issue.
Thank you,
Amos
------------------------------
Message: 3
Date: Fri, 24 Jun 2011 10:47:03 -0700
From: Delossantos_Roseann <Delossantos_Roseann <@t> Allergan.com>
Subject: RE: [Histonet] Histogel Problem
To: Amos Brooks <amosbrooks <@t> gmail.com>
Cc: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<6E159393AFE18142A83CE7C2475AD96D0916498C5A <@t> AGMAILCL100.allergan.com>
Content-Type: text/plain; charset="us-ascii"
Hi,
I've encountered this problem before in my previous lab. To reduce the Histogel from shrinking that badly, avoid putting the tissue at the very edge of the Histogel, space them out nicely in the middle, providing sufficient amount of extra Histogel between each tissue and surrounding them. A little bit of shrinking usually do not interfere with cutting as long as the whole thing is placed flat on the final paraffin block, the rest will stretch out on the water bath. I find sometimes the gel does not behave well in the water but as long as it is not on top of your tissue when placed on your slide, it should interfere much with staining.
Rose
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Amos Brooks
Sent: Friday, June 24, 2011 10:29 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Histogel Problem
Hi,
I have a problem with some blocks that were prepared with Histogel. I was
hoping someone else might have had a similar problem and figured it out. I
took a photo of the blocks that were mads and put them in a Picassa album
here:
https://picasaweb.google.com/lh/photo/APO3HsIMa2_jPOEs3QRGUjhz3qi22FNPb2i5JJnBCAk?feat=directlink
The long & short of it is that the blocs were prepared by the researcher
for me to process. They are mouse kidneys. Now it is entirely possible for
him to have goofed something up in preparing the Histogel blocks. I wasn't
there when he did it, but when I looked at them before processing, they all
looked fine. (Like the adjacent good one in the photo). When they were
processed they were placed in the VIP right next to each other. When I went
to embed them this morning all but one of the four looked fine. The one that
didn't come out well looked like the Histogel had shrunk up and shriveled
around the kidneys. I am sure this will be aweful to cut, and the researcher
is going to have a bird over it since this happened with another project
previously. I would like to have a decent explanation for him, so if anyone
knows what might have happened and has suggestions I would love to hear it
(yes vendors too are welcome to answer this of course!).
By the way, this was processed on a rather short cycle of 15-20 min per
station of graded ETOH from 70% to 100% with 3 xylene stations and 4
paraffin stations (45 min for these). It seems fine for everything else that
was on the processor. Just that one Histogel block was the issue.
Thank you,
Amos
_______________________________________________
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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------------------------------
Message: 4
Date: Fri, 24 Jun 2011 14:20:33 -0500
From: "Webb, Dorothy L" <Dorothy.L.Webb <@t> HealthPartners.Com>
Subject: [Histonet] bluing
To: "'histonet <@t> lists.utsouthwestern.edu'"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<65365F35C0F2EF4D846EC3CA73E49C43010F8BFEBD7D <@t> HPEMX3.HealthPartners.int>
Content-Type: text/plain; charset="us-ascii"
Looking to change my "bluing" step in the H&E process to obtain a bluer (less purple) hue to the nuclear detail. What is everyone using in their bluing step??
Thanks for all of your ideas!!
________________________________
This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited.
If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0
------------------------------
Message: 5
Date: Fri, 24 Jun 2011 12:35:18 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] bluing
To: "'histonet <@t> lists.utsouthwestern.edu'"
<histonet <@t> lists.utsouthwestern.edu>, Dorothy LWebb
<Dorothy.L.Webb <@t> HealthPartners.Com>
Message-ID: <201322.12289.qm <@t> web65716.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
There are several "bluing solutions" in the market, or you could use lithium carbonate at different concentrations until you find one of your liking.
Ren? J.
--- On Fri, 6/24/11, Webb, Dorothy L <Dorothy.L.Webb <@t> HealthPartners.Com> wrote:
From: Webb, Dorothy L <Dorothy.L.Webb <@t> HealthPartners.Com>
Subject: [Histonet] bluing
To: "'histonet <@t> lists.utsouthwestern.edu'" <histonet <@t> lists.utsouthwestern.edu>
Date: Friday, June 24, 2011, 3:20 PM
Looking to change my "bluing" step in the H&E process to obtain a bluer (less purple) hue to the nuclear detail.? What is everyone using in their bluing step??
Thanks for all of your ideas!!
? ________________________________
This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited.
If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0
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------------------------------
Message: 6
Date: Fri, 24 Jun 2011 15:33:12 -0500
From: "Harrison, Sandra C." <Sandra.Harrison3 <@t> va.gov>
Subject: RE: [Histonet] bluing
To: "Webb, Dorothy L" <Dorothy.L.Webb <@t> HealthPartners.Com>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<DB425E28065DA14FAA75A282959620AF0546BAEB <@t> VHAV23MSGA2.v23.med.va.gov>
Content-Type: text/plain; charset="us-ascii"
Hi Dorothy,
Try Richard Allan Bluing Reagent.
Here's what they say about their product: "It is a buffered product
that ensures the proper alkalinity (pH=8.0). Unlike ammonia and lithium
carbonate, RA's Bluing Reagent does not allow for the pH shift which can
affect the crispness of nuclear detail."
When I first began supervising this lab 5 years ago, they "made from
scratch" their own hematoxylin and eosin (not to mention buffered
formalin.) Unfortunately, the quality of the stain was very spotty and
caused the Pathologists a lot of problems. I switched us to the Richard
Allan 7211 Hematoxylin, which has beautiful, crisp nuclear detail. We
also went with the recommended Richard Allan Clarifier, Bluing and
Eosin, as well, so that we could produce a consistently high quality H&E
every time.
In 5 years, we've had very few complaints about the stain from the 8-10
Pathologists we've worked with, except for one occasion when there was
some isolated nuclear hazing. We did some detective work and determined
that the cause was due to a rack or two that had been placed in the oven
without properly removing the excess water or draining of the slides
before placing them in the oven.
Have a great week-end everybody. It's practically the 1st sunny day
we've had, here in Minneapolis, for the past 2 weeks and Saturday and
Sunday's forecast looks good, too!
Sandy Harrison
VA-Minneapolis
Supervisor, Anatomical and Surgical Pathology
612-467-2449
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Webb,
Dorothy L
Sent: Friday, June 24, 2011 2:21 PM
To: 'histonet <@t> lists.utsouthwestern.edu'
Subject: [Histonet] bluing
Looking to change my "bluing" step in the H&E process to obtain a bluer
(less purple) hue to the nuclear detail. What is everyone using in
their bluing step??
Thanks for all of your ideas!!
________________________________
This e-mail and any files transmitted with it are confidential and are
intended solely for the use of the individual or entity to whom they are
addressed. If you are not the intended recipient or the individual
responsible for delivering the e-mail to the intended recipient, please
be advised that you have received this e-mail in error and that any use,
dissemination, forwarding, printing, or copying of this e-mail is
strictly prohibited.
If you have received this e-mail in error, please immediately notify the
HealthPartners Support Center by telephone at (952) 967-6600. You will
be reimbursed for reasonable costs incurred in notifying us.
HealthPartners R001.0
_______________________________________________
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Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 7
Date: Fri, 24 Jun 2011 15:53:39 -0500
From: "Webb, Dorothy L" <Dorothy.L.Webb <@t> HealthPartners.Com>
Subject: [Histonet] blades
To: "'histonet <@t> lists.utsouthwestern.edu'"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<65365F35C0F2EF4D846EC3CA73E49C43010F8BFEBD81 <@t> HPEMX3.HealthPartners.int>
Content-Type: text/plain; charset="us-ascii"
Trying to clean up some things hanging out there in our lab and wondering what everyone does with a blade that has been used minimally and tech done for the day with the microtome. Where do you store that blade for use tomorrow or do you toss and not worry about the cost involved? I do not like them sitting on top of the microtome. Any good ideas?? Thanks, as always!
________________________________
This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited.
If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0
------------------------------
Message: 8
Date: Fri, 24 Jun 2011 17:16:06 -0400
From: Sean McBride <smcbride <@t> andrew.cmu.edu>
Subject: Re: [Histonet] blades
To: "Webb, Dorothy L" <Dorothy.L.Webb <@t> HealthPartners.Com>
Cc: "'histonet <@t> lists.utsouthwestern.edu'"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <0FA5973D-A2CA-4364-BD5E-A477F302BD8D <@t> andrew.cmu.edu>
Content-Type: text/plain; charset=us-ascii
Dorothy,
I put ours in a 15 mL centrifuge tube with a cap & sit it on the base of the microtome for the next use, that way, no one gets cut & the blade is able to be used to the fullest of it's potential. :-)
Best regards,
~Sean McBride
Scientific Specialist
Bone Tissue Engineering Center
Carnegie Mellon Research Institute
Suite 4311
700 Technology Drive
Pittsburgh, PA 15219-3124
412-268-8275 (o)
412-915-1683 (m)
412-268-8275 (fax)
smcbride <@t> andrew.cmu.edu
On Jun 24, 2011, at 4:53 PM, Webb, Dorothy L wrote:
> Trying to clean up some things hanging out there in our lab and wondering what everyone does with a blade that has been used minimally and tech done for the day with the microtome. Where do you store that blade for use tomorrow or do you toss and not worry about the cost involved? I do not like them sitting on top of the microtome. Any good ideas?? Thanks, as always!
>
>
>
> ________________________________
> This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited.
>
> If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
------------------------------
Message: 9
Date: Fri, 24 Jun 2011 17:10:30 -0400
From: Esther C Peters <epeters2 <@t> gmu.edu>
Subject: Re: [Histonet] blades
To: "Webb, Dorothy L" <Dorothy.L.Webb <@t> HealthPartners.Com>
Cc: "'histonet <@t> lists.utsouthwestern.edu'"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <fa3e9f5c1d7b4.4e04c506 <@t> gmu.edu>
Content-Type: text/plain; charset=us-ascii
We put ours in a small slide box (plastic or styrofoam, 5-25 slides) that is clearly marked "Microtome Blades for Facing Blocks" to be used another day.
Esther C. Peters, Ph.D.
Assistant Professor
Department of Environmental Science & Policy
Biology Program/Medical Technology Coordinator
George Mason University
4400 University Drive, MSN 5F2
Fairfax, VA 22030-4444
Office: David King Hall 3057
Phone: 703-993-3462
Fax: 703-993-1066
epeters2 <@t> gmu.edu
----- Original Message -----
From: "Webb, Dorothy L" <Dorothy.L.Webb <@t> HealthPartners.Com>
Date: Friday, June 24, 2011 4:53 pm
Subject: [Histonet] blades
> Trying to clean up some things hanging out there in our lab and
> wondering what everyone does with a blade that has been used
> minimally and tech done for the day with the microtome. Where do
> you store that blade for use tomorrow or do you toss and not worry
> about the cost involved? I do not like them sitting on top of the
> microtome. Any good ideas?? Thanks, as always!
>
>
>
> ________________________________
> This e-mail and any files transmitted with it are confidential and
> are intended solely for the use of the individual or entity to
> whom they are addressed. If you are not the intended recipient or
> the individual responsible for delivering the e-mail to the
> intended recipient, please be advised that you have received this
> e-mail in error and that any use, dissemination, forwarding,
> printing, or copying of this e-mail is strictly prohibited.
>
> If you have received this e-mail in error, please immediately
> notify the HealthPartners Support Center by telephone at (952) 967-
> 6600. You will be reimbursed for reasonable costs incurred in
> notifying us. HealthPartners R001.0
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
------------------------------
Message: 10
Date: Fri, 24 Jun 2011 17:31:33 -0400
From: Victoria Baker <bakevictoria <@t> gmail.com>
Subject: Re: [Histonet] blades
To: Esther C Peters <epeters2 <@t> gmu.edu>
Cc: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>, "Webb, Dorothy L"
<Dorothy.L.Webb <@t> healthpartners.com>
Message-ID: <BANLkTinw2CqTSXPyuyq=UDxAZpHwYemrKQ <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1
I use the cardboards that come in a box of slides. A small piece of tape on
the open side and mark it used. I just always make sure I have the blade
edge facing the folded part. I know some who will tape this folded
board it to the side of their microtome and use it as a trimming blade
holder.
Vikki
On Fri, Jun 24, 2011 at 5:10 PM, Esther C Peters <epeters2 <@t> gmu.edu> wrote:
> We put ours in a small slide box (plastic or styrofoam, 5-25 slides) that
> is clearly marked "Microtome Blades for Facing Blocks" to be used another
> day.
>
> Esther C. Peters, Ph.D.
> Assistant Professor
> Department of Environmental Science & Policy
> Biology Program/Medical Technology Coordinator
> George Mason University
> 4400 University Drive, MSN 5F2
> Fairfax, VA 22030-4444
> Office: David King Hall 3057
> Phone: 703-993-3462
> Fax: 703-993-1066
> epeters2 <@t> gmu.edu
>
> ----- Original Message -----
> From: "Webb, Dorothy L" <Dorothy.L.Webb <@t> HealthPartners.Com>
> Date: Friday, June 24, 2011 4:53 pm
> Subject: [Histonet] blades
>
> > Trying to clean up some things hanging out there in our lab and
> > wondering what everyone does with a blade that has been used
> > minimally and tech done for the day with the microtome. Where do
> > you store that blade for use tomorrow or do you toss and not worry
> > about the cost involved? I do not like them sitting on top of the
> > microtome. Any good ideas?? Thanks, as always!
> >
> >
> >
> > ________________________________
> > This e-mail and any files transmitted with it are confidential and
> > are intended solely for the use of the individual or entity to
> > whom they are addressed. If you are not the intended recipient or
> > the individual responsible for delivering the e-mail to the
> > intended recipient, please be advised that you have received this
> > e-mail in error and that any use, dissemination, forwarding,
> > printing, or copying of this e-mail is strictly prohibited.
> >
> > If you have received this e-mail in error, please immediately
> > notify the HealthPartners Support Center by telephone at (952) 967-
> > 6600. You will be reimbursed for reasonable costs incurred in
> > notifying us. HealthPartners R001.0
> > _______________________________________________
> > 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
>
------------------------------
Message: 11
Date: Fri, 24 Jun 2011 14:50:43 -0700
From: Jennifer MacDonald <jmacdonald <@t> mtsac.edu>
Subject: Re: [Histonet] blades
To: "Webb, Dorothy L" <Dorothy.L.Webb <@t> HealthPartners.Com>
Cc: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <9A2E9444-36FF-418C-A794-AB6531E953AD <@t> mtsac.edu>
Content-Type: text/plain; charset=us-ascii
We use plastic 5 slide mailers
Sent from my iPhone
On Jun 24, 2011, at 1:53 PM, "Webb, Dorothy L" <Dorothy.L.Webb <@t> HealthPartners.Com> wrote:
> Trying to clean up some things hanging out there in our lab and wondering what everyone does with a blade that has been used minimally and tech done for the day with the microtome. Where do you store that blade for use tomorrow or do you toss and not worry about the cost involved? I do not like them sitting on top of the microtome. Any good ideas?? Thanks, as always!
>
>
>
> ________________________________
> This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited.
>
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------------------------------
Message: 12
Date: Fri, 24 Jun 2011 16:56:32 -0700
From: Eric Velazquez <ervelazquez <@t> gmail.com>
Subject: [Histonet] HT Position - Irvine, CA
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <BANLkTikBB4HqvgM=UTtRrGMC9B2sbZ3khQ <@t> mail.gmail.com>
Content-Type: text/plain; charset=windows-1252
Hi Histonet,
We are currently looking for a full-time Histotechnician (preferably HT
certified) with experience in IHC. Please read the description below and if
you're interested please submit your resume via e-mail to: *
eric.velazquez <@t> agendia.com*.
*ESSENTIAL DUTIES AND RESPONSIBILITIES*
- Performs histology aspects of the lab, which includes cutting
tissue(frozen, FFPE),
manually staining sections for H&E imaging, IHC staining, loading slides
on ScanScope for imaging and scoring by pathologist
- Maintains laboratory logs so that current information is complete and
readily available to staff, including clinical log sheet, ScanScope
identification
- Performs all aspects of lab support to ensure timely completion of
samples. Reports any discrepancies to the Director Of Laboratory
Operations.
- Performs various maintenance and/or housekeeping tasks to keep
laboratory clean, and ensure ease of use.
- Cleans and fills water baths, bulk reagents, dump waste containers
- Reports any malfunctions of equipment to the Director Of Laboratory
Operations.
- Washes all laboratory dishes to ensure a clean supply when needed.
- Performs data entry tasks in the laboratory and office when needed.
- Follows all Agendia, Inc.?s health and safety policies and procedures
- Performs other related duties as required or assigned
Thank you for your interest.
-Eric Velazquez
Agendia Inc.
------------------------------
Message: 13
Date: Sat, 25 Jun 2011 10:10:17 -0400
From: "Lee & Peggy Wenk" <lpwenk <@t> sbcglobal.net>
Subject: Re: [Histonet] HT Position - Irvine, CA
To: "Eric Velazquez" <ervelazquez <@t> gmail.com>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <803AB1A1F69C40708BC7795115D1CFB7 <@t> HP2010>
Content-Type: text/plain; format=flowed; charset="Windows-1252";
reply-type=original
Where are you? City, State, Country, Lab, etc. Info, please.
-----Original Message-----
From: Eric Velazquez
Sent: Friday, June 24, 2011 7:56 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] HT Position - Irvine, CA
Hi Histonet,
We are currently looking for a full-time Histotechnician (preferably HT
certified) with experience in IHC. Please read the description below and if
you're interested please submit your resume via e-mail to: *
eric.velazquez <@t> agendia.com*.
*ESSENTIAL DUTIES AND RESPONSIBILITIES*
- Performs histology aspects of the lab, which includes cutting
tissue(frozen, FFPE),
manually staining sections for H&E imaging, IHC staining, loading slides
on ScanScope for imaging and scoring by pathologist
- Maintains laboratory logs so that current information is complete and
readily available to staff, including clinical log sheet, ScanScope
identification
- Performs all aspects of lab support to ensure timely completion of
samples. Reports any discrepancies to the Director Of Laboratory
Operations.
- Performs various maintenance and/or housekeeping tasks to keep
laboratory clean, and ensure ease of use.
- Cleans and fills water baths, bulk reagents, dump waste containers
- Reports any malfunctions of equipment to the Director Of Laboratory
Operations.
- Washes all laboratory dishes to ensure a clean supply when needed.
- Performs data entry tasks in the laboratory and office when needed.
- Follows all Agendia, Inc.?s health and safety policies and procedures
- Performs other related duties as required or assigned
Thank you for your interest.
-Eric Velazquez
Agendia Inc.
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 14
Date: Sat, 25 Jun 2011 07:16:32 -0700 (PDT)
From: Aurea Marquez <amarquez29 <@t> yahoo.com>
Subject: [Histonet] Contents of Histonet digest
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <462293.86417.qm <@t> web120511.mail.ne1.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
?
Aurea Griselle Marquez
________________________________
From: "histonet-request <@t> lists.utsouthwestern.edu"
<histonet-request <@t> lists.utsouthwestern.edu>
To: histonet <@t> lists.utsouthwestern.edu
Sent: Fri, June 24, 2011 1:01:21 PM
Subject: Histonet Digest, Vol 91, Issue 33
Send Histonet mailing list submissions to
??? histonet <@t> lists.utsouthwestern.edu
To subscribe or unsubscribe via the World Wide Web, visit
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or, via email, send a message with subject or body 'help' to
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You can reach the person managing the list at
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When replying, please edit your Subject line so it is more specific
than "Re: Contents of Histonet digest..."
Today's Topics:
? 1. Formula R paraffin (Clare Thornton)
? 2. Re: Formula R paraffin (Rene J Buesa)
? 3. HistoGel (Dessoye, Michael J)
? 4. (no subject) (Kendra)
----------------------------------------------------------------------
Message: 1
Date: Fri, 24 Jun 2011 12:08:02 -0400
From: Clare Thornton <CThornton <@t> dahlchase.com>
Subject: [Histonet] Formula R paraffin
To: "'Histonet <@t> lists.utsouthwestern.edu'"
??? <Histonet <@t> lists.utsouthwestern.edu>
Message-ID:
??? <C9D78FFC9D668B4CBEA4405F84697504F8144E556A <@t> iris.dahlchase.net>
Content-Type: text/plain; charset="us-ascii"
Does anyone have any experience with Formula R paraffin, from Leica, good or
bad?? We currently use Paraplast X-tra but occasionally have troubles with
compression, and we heard Formula R is harder and less likely to compress.
Thanks!
Clare J. Thornton, HTL(ASCP), QIHC
Assistant Histology Supervisor
Dahl-Chase Diagnostic Services
417 State Street, Suite 540
Bangor, ME 04401
cthornton <@t> dahlchase.com
------------------------------
Message: 2
Date: Fri, 24 Jun 2011 09:21:35 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Formula R paraffin
To: "'Histonet <@t> lists.utsouthwestern.edu'"
??? <Histonet <@t> lists.utsouthwestern.edu>, ??? Clare Thornton
??? <CThornton <@t> dahlchase.com>
Message-ID: <851084.79705.qm <@t> web65712.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
If sometimes you are experiencing compression problems it could be because your
paraffin has some xylene contamination, or your blocks are not cold enough when
cutting, or you are using a paraffin of lower melting point as needed for the
tissue you regularly process.
Paraplast X-tra always worked fine for me. Check with them for a harder paraffin
instead of changing to a different paraffin altogether.
Ren? J.
--- On Fri, 6/24/11, Clare Thornton <CThornton <@t> dahlchase.com> wrote:
From: Clare Thornton <CThornton <@t> dahlchase.com>
Subject: [Histonet] Formula R paraffin
To: "'Histonet <@t> lists.utsouthwestern.edu'" <Histonet <@t> lists.utsouthwestern.edu>
Date: Friday, June 24, 2011, 12:08 PM
Does anyone have any experience with Formula R paraffin, from Leica, good or
bad?? We currently use Paraplast X-tra but occasionally have troubles with
compression, and we heard Formula R is harder and less likely to compress.
Thanks!
Clare J. Thornton, HTL(ASCP), QIHC
Assistant Histology Supervisor
Dahl-Chase Diagnostic Services
417 State Street, Suite 540
Bangor, ME 04401
cthornton <@t> dahlchase.com
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 3
Date: Fri, 24 Jun 2011 12:26:46 -0400
From: "Dessoye, Michael J" <mjdessoye <@t> wvhcs.org>
Subject: [Histonet] HistoGel
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
??? <E2547E1CD0EE324488A2940994571EFA0401F3AE <@t> WVHCS-EXCHANGE.wvhcs.com>
Content-Type: text/plain;??? charset="iso-8859-1"
Hello,
Does anyone out there have any experience with HistoGel?? It's Richard
Allan/Thermo Fisher.? They claim that you can "embed" scant tissues in the gel
and then process, embed, and cut as usual.? Just wondering how it works in the
real world....
Michael J. Dessoye, M.S. | Histology Supervisor | Wyoming Valley Health Care
System | mjdessoye <@t> wvhcs.org <mailto:mjdessoye <@t> wvhcs.org>? |
575 N. River Street | Wilkes Barre, PA 18764 | Tel: 570-552-1485 | Fax:
570-552-1526
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------------------------------
Message: 4
Date: Fri, 24 Jun 2011 11:28:55 -0500
From: Kendra <shultz11 <@t> cox.net>
Subject: [Histonet] (no subject)
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <723pa4ptd019lmddnpqhrxwt.1308932935185 <@t> email.android.com>
Content-Type: text/plain; charset=utf-8
------------------------------
Message: 15
Date: Sat, 25 Jun 2011 10:26:23 -0400
From: "Lee & Peggy Wenk" <lpwenk <@t> sbcglobal.net>
Subject: Re: [Histonet] bluing
To: "Webb, Dorothy L" <Dorothy.L.Webb <@t> HealthPartners.Com>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <2B7338AFC93A4F91AD215CDC80E1B201 <@t> HP2010>
Content-Type: text/plain; format=flowed; charset="iso-8859-1";
reply-type=original
It might just need longer time in the bluing agent.
Hematoxylin solutions stain the nuclei reddish (look at your slide right
after coming out of hematoxylin - tissue will look reddish). The alkaline
solutions that are used for "bluing" remove a H+ group on the aluminum
hematein (that's the staining chemical of the hematoxylin solution), and
change it to a -OH group. This changes the aluminum hematein from a reddish
color to a blue color.
The more alkaline (higher pH) the bluing agent, the faster this reaction and
color change, and the less time is needed in the bluing agent. The lower the
alkalinity (not as high pH but still in the alkaline range) the bluing
agent, the slower the reaction and color change, and more time is needed in
the bluing agent. Dilute ammonia water usually only takes a few seconds, as
it's pH is usually high (pH 10). Tap water can have a pH of around 7, and
may take 5-10 minutes to "blue". If the tap water is more acidic (pH 5 or
below), the slides may not "blue".
Now, we have to get all the nuclei from the reddish color to the bluish
color. If the slide is not in the bluing agent for enough time (for the
type/pH of bluing agent), then some of the nuclei change to blue, while some
still remain reddish (or within a single nucleus, some of the DNA has
changed to blue, some remains reddish), hence a more purple color.
So, the easiest thing to try right now is to stain 2 slides from the same
block (serial sections) in your hematoxylin for the same time, put one in
the bluing agent for the usual time, put the other in the alkaline solution
for extended time, and see if the nuclei on the second slide are now more
blue than the first slide.
Let us know.
Peggy A. Wenk, HTL(ASCP)SLS
Beaumont Hospital
Royal Oak, MI 48073
-----Original Message-----
From: Webb, Dorothy L
Sent: Friday, June 24, 2011 3:20 PM
To: 'histonet <@t> lists.utsouthwestern.edu'
Subject: [Histonet] bluing
Looking to change my "bluing" step in the H&E process to obtain a bluer
(less purple) hue to the nuclear detail. What is everyone using in their
bluing step??
Thanks for all of your ideas!!
________________________________
This e-mail and any files transmitted with it are confidential and are
intended solely for the use of the individual or entity to whom they are
addressed. If you are not the intended recipient or the individual
responsible for delivering the e-mail to the intended recipient, please be
advised that you have received this e-mail in error and that any use,
dissemination, forwarding, printing, or copying of this e-mail is strictly
prohibited.
If you have received this e-mail in error, please immediately notify the
HealthPartners Support Center by telephone at (952) 967-6600. You will be
reimbursed for reasonable costs incurred in notifying us. HealthPartners
R001.0
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 16
Date: Sat, 25 Jun 2011 12:00:44 -0400
From: Sheila Adey <sadey <@t> hotmail.ca>
Subject: [Histonet] Leica Bond for IHCs
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <BAY154-w67E5E7B918DAB1D2C341CC6550 <@t> phx.gbl>
Content-Type: text/plain; charset="iso-8859-1"
Hello netters:
Looking for opinions on the Leica Bond immuno stainer please.
Thanks.
Sheila
------------------------------
Message: 17
Date: Sat, 25 Jun 2011 10:47:15 -0600
From: "gayle callis" <gayle.callis <@t> bresnan.net>
Subject: [Histonet] RE: Bluing
To: "Histonet" <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <000101cc3357$8b945ff0$a2bd1fd0$@callis <@t> bresnan.net>
Content-Type: text/plain; charset="us-ascii"
Dear All,
We use commercial bluing reagents for convenience e.g. Richard Allan Bluing
Reagent ready to use and have used Scotts Tap Water Substitute Concentrate
diluted before use. We do not use ammonia water anymore as the high pH can
cause section lift off from slide. Here is recipe for Scotts.
SCOTT'S TAP WATER SUBSTITUTE (You can make this up as a 10X concentrate if
you wish. Store in house concentrate at 4C or growth will occur)
Sodium bicarbonate 2 g
Magnesium sulfate (anhydrous) 20 g
Tap Water 1 L
Peggy presented a good explanation of bluing. We do not use tap water
anymore due to incorrect pH. One of the failures for complete bluing is
not changing the bluing reagent daily with large volumes of slides. Lower
number of slides, one might be able to get by with fewer changes of bluing.
Tap water rinses dilutes the reagent and can change the pH, which should be
pH 8. If there is a large volume of slides, anything over 75 or more, we
change the bluing reagent daily. It is important to remember that Clarifiers
e.g. acetic acid reagents used with progressive hematoxylins (Richard Allan
Hematoxylin 1, Gill 1,2,3) will suffer from tap water rinse dilution too. We
also change those, and if Clarifier looks rather yellow, then you have
probably exhausted the acetic acid and its effectiveness. Jerry Fredenburgh
has given excellent workshops on hematoxylin staining in the past and passed
on many of these facts. Consequently, we have no clarifying or bluing
problems.
Fredenburgh recommendations for Richard Allan Hematoxylin 1 staining
procedure that we use:
Hematoxylin 1 - 1.5 min, longer if needed
Tap water rinse-1 minute
Clarifier - 1 minute
Tap water rinse - 1 minute
Bluing - 1 minute
Tap water rinse - 1 minute
Remember, static or non-running water rinse stations are contaminated with
carryover hematoxylin, clarifier and/or bluing reagent. If you do NOT
remove the bluing reagent with a good rinse, then eosin staining is affected
by cations carried over from the bluing reagent. We also have an alcohol
step (matching % of alcohol in eosin) before going into this stain. This
helps remove the cations plus prepares the section for eosin staining.
Perform good rinses and change static water rinse stations after a run, or
frequently during a day of heavy staining. The second and third racks of
slides on a stainer should have the same tap water rinse conditions as the
first rack. Yes, extra work but you need to ensure consistent staining
results of H&E of ALL sections- the universal workhorse stain we depend on
so heavily.
Years ago, Richard Allan had an excellent manual for H&E staining guidelines
on how to perform and adjust your H&E staining. Sadly the manual is no
longer published but the good news is ...... I saved my copy, and scanned
it to pdf if anyone wants it.
Gayle M. Callis
HTL/HT/MT(ASCP)
Bozeman MT
****************************************************************************
*****************************************************************
Peggy Wenk wrote:
It might just need longer time in the bluing agent.
Hematoxylin solutions stain the nuclei reddish (look at your slide right
after coming out of hematoxylin - tissue will look reddish). The alkaline
solutions that are used for "bluing" remove a H+ group on the aluminum
hematein (that's the staining chemical of the hematoxylin solution), and
change it to a -OH group. This changes the aluminum hematein from a reddish
color to a blue color.
The more alkaline (higher pH) the bluing agent, the faster this reaction and
color change, and the less time is needed in the bluing agent. The lower the
alkalinity (not as high pH but still in the alkaline range) the bluing
agent, the slower the reaction and color change, and more time is needed in
the bluing agent. Dilute ammonia water usually only takes a few seconds, as
it's pH is usually high (pH 10). Tap water can have a pH of around 7, and
may take 5-10 minutes to "blue". If the tap water is more acidic (pH 5 or
below), the slides may not "blue".
Now, we have to get all the nuclei from the reddish color to the bluish
color. If the slide is not in the bluing agent for enough time (for the
type/pH of bluing agent), then some of the nuclei change to blue, while some
still remain reddish (or within a single nucleus, some of the DNA has
changed to blue, some remains reddish), hence a more purple color.
So, the easiest thing to try right now is to stain 2 slides from the same
block (serial sections) in your hematoxylin for the same time, put one in
the bluing agent for the usual time, put the other in the alkaline solution
for extended time, and see if the nuclei on the second slide are now more
blue than the first slide.
Let us know.
Peggy A. Wenk, HTL(ASCP)SLS
Beaumont Hospital
Royal Oak, MI 48073
-----Original Message-----
From: Webb, Dorothy L
Sent: Friday, June 24, 2011 3:20 PM
To: 'histonet <http://lists.utsouthwestern.edu/mailman/listinfo/histonet>
<@t> lists.utsouthwestern.edu'
Subject: [Histonet] bluing
Looking to change my "bluing" step in the H&E process to obtain a bluer
(less purple) hue to the nuclear detail. What is everyone using in their
bluing step??
Thanks for all of your ideas!!
------------------------------
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
End of Histonet Digest, Vol 91, Issue 34
****************************************
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