[Histonet] RE: Histonet Digest, Vol 129, Issue 25
Algeo, Lacie A
Lacie.Algeo <@t> providence.org
Mon Aug 18 11:18:18 CDT 2014
I hope I am responding to the correct location :)
Brandy....a few questions about your tissue falling off your slides. What kind of tissue? What kind of slides? What kind of stainer?
Thanks,
Lacie
Lacie Algeo, HTL (ASCP) MBCM
Histology Supervisor
Providence Sacred Heart Medical Center Laboratory
101 W 8th Avenue
L-2
Spokane, WA 99204
509-474-4418
FAX 509-474-2052
lacie.algeo <@t> providence.org
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Subject: Histonet Digest, Vol 129, Issue 25
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Today's Topics:
1. antibody suggestions CD8, FAP, Cd11b, CD11c (Mesru T)
2. RE: Tissue falling off (Terri Braud)
3. Re: RE: Glassware Cleaning again (darkdaym <@t> comcast.net)
4. Re: antibody suggestions CD8, FAP, Cd11b, CD11c (gayle callis)
5. Re: RE: Glassware Cleaning again (E. Wayne Johnson)
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Message: 1
Date: Fri, 15 Aug 2014 13:24:29 -0400
From: Mesru T <turkekul <@t> gmail.com>
Subject: [Histonet] antibody suggestions CD8, FAP, Cd11b, CD11c
To: histonet <@t> lists.utsouthwestern.edu
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Dear Histonetters,
I need your help to find good antibodies for IHC on mouse and human FFPE tissues like normal tissues and tumors. Please share any information you think might be helpful.
CD8 for IHC on mouse FFPE spleen or other lymphoid organs.
FAP (Fibroblast Activated protein) for IHC on mouse and human FFPE tumors.
CD11b for IHC on mouse FFPE spleen or other lymphoid organs..
CD11c for IHC on mouse FFPE spleen or other lymphoid organs.
Thanks in advance for all your help.
Regards,
Mesru
------------------------------
Message: 2
Date: Fri, 15 Aug 2014 13:52:09 -0400
From: "Terri Braud" <tbraud <@t> holyredeemer.com>
Subject: [Histonet] RE: Tissue falling off
To: <histonet <@t> lists.utsouthwestern.edu>
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<BFBF7C6B9627B947B74CBEBD45CE368B14048B56 <@t> hrex-svr.holyredeemer.local>
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Hi - Just a couple of suggestions -
I've seen this periodically crop up during the years. There are a couple of things to consider:
Water baths are completely clean - after dumpling water and removing paraffin debris with whatever you use - wipe out the bath with clean absolute alcohol.
Be careful where you spray - make sure that if you use anything like Paraguard, or similar stuff that keeps paraffin from sticking, that it is used sparingly, or preferably, not at all. We had a situation where we had a trial sample that someone was using to clean her microtome and forceps and the stuff got everywhere and tissues started falling off.
Your water in the waterbaths is clean - Fresh DI water. Once we had a contaminant in our DI water, and the patient tissues slid right off our slides. We use clean slides, good quality water and super clean waterbaths.
Is your heater up and running? - Our slides are stained on a stainer with built in slide dryers. We used to have a problem with the first rack of patient slides' tissue falling off because the slide dryer was not hot enough for our short dry cycle. We now send a rack through first, which turns on the heaters, then load the patient rack.
Hope this gives you a few ideas
Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Holy Redeemer Hospital Laboratory
1648 Huntingdon Pike
Meadowbrook, PA 19046
Ph: 215-938-3676
Fax: 215-938-3874
7. tissue falling off slides (Burnett, Brandy)
From: "Burnett, Brandy" <bburnett <@t> CapeCodHealth.org>
Subject: [Histonet] tissue falling off slides
Our lab has encountered an issue with tissue falling off of the slides.
It is mostly the patient tissue
that is falling off. Some of the Techs are using control slides that have been pre-cut and I am wondering if this might be part of the problem. Any information would be very much appreciated.
Thanks Again,
Brandy Burnett, HTL
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Message: 3
Date: Fri, 15 Aug 2014 13:06:52 -0500
From: darkdaym <@t> comcast.net
Subject: Re: [Histonet] RE: Glassware Cleaning again
To: histonet <@t> lists.utsouthwestern.edu
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Brian, et.al.,
It's not a typo or and an error. It's one might call a variant spelling. The Merck Index lists Bromcresol Purple, CAS Number 115-40-2.
Below is what Sigma lists with the same CAS number. I've noticed the prefix Brom or Bromo on the name of several dyes. Either way, it's the same thing.
*114375 * *Sigma-Aldrich *
Bromocresol Purple
BioReagent, suitable for indicator, Dye content 90 %
Synonym: *5,5?-Dibromo-/o/-cresolsulfonphthalein, Bromcresol purple sultone form
*
On 8/15/2014 11:35 AM, Cooper, Brian wrote:
> I noticed the discrepancy in spelling too. I looked online for like 30 minutes, and couldn't find anything called "Bromcresol." Found a lot of vendors selling Bromocresol Purple (and Green for that matter). Best I can figure is that this is a typo on CAP's checklist (that's been there for several years now).
>
> Thanks,
>
> Brian D. Cooper, HT (ASCP)CM | Histology Supervisor
> Department of Pathology and Laboratory Medicine
> Children's Hospital Los Angeles
> 4650 Sunset Blvd MS#43- Los Angeles, CA 90027
> bcooper <@t> chla.usc.edu
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Laurie Colbert
> Sent: Friday, August 15, 2014 9:13 AM
> To: Histonet Post (histonet <@t> lists.utsouthwestern.edu)
> Subject: [Histonet] Glassware Cleaning again
>
> Is bromocresol purple the same as bromcresol purple? The CAP question regarding glassware cleaning refers to bromcresol purple, but I ordered a powder and it is labeled as bromocresol purple.
>
> Laurie Colbert, HT (ASCP)
> Histology Supervisor
> PATH MD
> 8158 Beverly Blvd.
> Los Angeles, CA 90048
> (323) 648-3214 direct
> (424) 245-7284 main lab
>
> The information contained in this transmission may contain privileged and confidential information, including patient information protected by federal and state privacy laws. It is intended only for the use of the person(s) named above. If you are not the intended recipient, you are hereby notified that any review, dissemination, distribution, or duplication of this communication is strictly prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message.
>
>
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------------------------------
Message: 4
Date: Fri, 15 Aug 2014 15:10:27 -0600
From: "gayle callis" <gayle.callis <@t> bresnan.net>
Subject: [Histonet] Re: antibody suggestions CD8, FAP, Cd11b, CD11c
To: <histonet <@t> lists.utsouthwestern.edu>
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You Wrote:
I need your help to find good antibodies for IHC on mouse and human FFPE
tissues like normal tissues and tumors. Please share any information you
think might be helpful.
CD8 for IHC on mouse FFPE spleen or other lymphoid organs.
FAP (Fibroblast Activated protein) for IHC on mouse and human FFPE tumors.
CD11b for IHC on mouse FFPE spleen or other lymphoid organs..
CD11c for IHC on mouse FFPE spleen or other lymphoid organs.
Thanks in advance for all your help.
Regards,
Mesru
____________________________________________________________________________
___________________________________________
I can answer some of the murine CD marker questions.
CD8 on FFPE mouse tissues will never work. These markers are too strongly
cross linked by the aldehyde based fixation, and PFA is not an alternative.
You can try HIER and Enzyme digestions until you are blue in the face, and
the staining will NOT work. This will also be true of CD11c dendritic cell
Clone HL3, and CD11b Mac1.
This means you will have to use fresh tissue frozen sections, acetone fixed
for any staining success. We use an acetone/absolute ethanol mixture for
fixation of murine tissue for all our CD marker staining, especially when
you need to do all these CD markers on the same sample.
As for positive tissue controls,
CD8 Rat antiMouse monoclonal - normal spleen
CD11c - Peyers patches from small intestine, lymph node and probably spleen
will work.
CD11b, Mac1 - we stimulated mouse to produce macrophages in Peyers patches
located on small intestine. Stimulation was oral inoculation with an
attenuated bacteria.
FAP (Fibroblast Activated protein) for IHC on mouse and human FFPE tumors.
Go to ABCAM, and look at cross reactivity for this polyclonal antibody. It
works on FFPE mouse, human and two other species.
I can't provide an answer for FFPE human tissues with any of these CD
markers.
I strongly suggest you go to BD Bioscience and do a search for the CD
markers, especially the mouse that will be either rat and Armenian Hamster
monoclonals. Read the Technical data sheets and see what applications will
work or not in terms of IHC. We have used BD Bioscience, and eBioscience for
our murine CD marker with great success but you have to be aware that CD8
has more than one clone, some working better on solvent fixed murine tissues
than others. This is also true of CD11c. ABCAM has an excellent website
for these antibodies, but compare prices.
There is another fixative that is formalin free, the Becksteads IHC zinc
fixative and also PLP i.e. paraformaldehyde-lysine-Periodate. PLP may allow
staining of these aldehyde compromised CD markers since the lysine helps get
rid of free aldehydes, along with the periodate and is, however, a fixative
that is bit tedious to use since it has to be made up fresh every time you
use it. Beckstead originally developed IHC zinc fixative in order to do IHC
on human tumors, lymphomas if I recall. I have original and other
publications for both fixatives showing success with murine CD markers and
other species. The IHC zinc fixative allows paraffin embedding and IHC
staining of these markers, (also a publication by Nitta et al)
Our lab used fresh tissue frozen sections, acetone/alcohol fixed exclusively
for all CD marker IHC and/or single, double and triple immunofluroescence
protocols.
I will be happy to provide acetone/alcohol fixative method and alternative
acetone fixations that work with murine CD markers. You cannot use
acetone/alcohol for human CD8 or CD4 as the alcohol ruins the antigen -
however, FFPE probably will work with human markers. I can also provide
publications, information about PLP, Becksteads IHC zinc fixative. PLP
recipes can be found on the internet but I have one that is excellent for
larger volumes of this fixative. Biocare has information on this fixative
with their IHC products too.
Gayle M. Callis
HTL/HT/MT(ASCP)
------------------------------
Message: 5
Date: Sat, 16 Aug 2014 21:05:06 +0800
From: "E. Wayne Johnson" <ewj <@t> pigsqq.org>
Subject: Re: [Histonet] RE: Glassware Cleaning again
To: "Cooper, Brian" <bcooper <@t> chla.usc.edu>, Laurie Colbert
<lcolbert <@t> pathmdlabs.com>, "Histonet Post
(histonet <@t> lists.utsouthwestern.edu)"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <53EF5702.1080808 <@t> pigsqq.org>
Content-Type: text/plain; charset=ISO-8859-1; format=flowed
Bromcresol means Bromocresol.
Faint not, but I certainly can't say you won't dye if you leave out the O.
The "o" is subject to elision due to its difficulty in pronounciation.
Such an occurrence of elision between 2 consonants is called syncope.
In English writing an elided vowel is often replaced with an apostrophe to
indicate the elision and perhaps demonstrate the dialect of the speaker.
Colloquial dialects in writing are too informal for stuffy scientific
and medical types, and indeed
special meaning is liable to be construed to the presence of a spurious
punctuation in the name of a chemical.
We don't see {brom'cresol} in
the lab {purple, green} but we can hear of an existence
where it lives happily under other nomenclature.
So "bromcresol" means "bromocresol".
And as in American politics,
the removal of an "O" might seem to make a difference visually
and even might be comfortable to some,
but actually would amount absolutely nothing in terms of what is
being represented, or the dying that is going on.
On 8/16/2014 12:35 AM, Cooper, Brian wrote:
> I noticed the discrepancy in spelling too. I looked online for like 30 minutes, and couldn't find anything called "Bromcresol." Found a lot of vendors selling Bromocresol Purple (and Green for that matter). Best I can figure is that this is a typo on CAP's checklist (that's been there for several years now).
>
> Thanks,
>
> Brian D. Cooper, HT (ASCP)CM | Histology Supervisor
> Department of Pathology and Laboratory Medicine
> Children's Hospital Los Angeles
> 4650 Sunset Blvd MS#43- Los Angeles, CA 90027
> bcooper <@t> chla.usc.edu
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Laurie Colbert
> Sent: Friday, August 15, 2014 9:13 AM
> To: Histonet Post (histonet <@t> lists.utsouthwestern.edu)
> Subject: [Histonet] Glassware Cleaning again
>
> Is bromocresol purple the same as bromcresol purple? The CAP question regarding glassware cleaning refers to bromcresol purple, but I ordered a powder and it is labeled as bromocresol purple.
>
> Laurie Colbert, HT (ASCP)
> Histology Supervisor
> PATH MD
> 8158 Beverly Blvd.
> Los Angeles, CA 90048
> (323) 648-3214 direct
> (424) 245-7284 main lab
>
> The information contained in this transmission may contain privileged and confidential information, including patient information protected by federal and state privacy laws. It is intended only for the use of the person(s) named above. If you are not the intended recipient, you are hereby notified that any review, dissemination, distribution, or duplication of this communication is strictly prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message.
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ---------------------------------------------------------------------
> CONFIDENTIALITY NOTICE: This e-mail message, including any attachments,
> is for the sole use of the intended recipient(s) and may contain confidential
> or legally privileged information. Any unauthorized review, use, disclosure
> or distribution is prohibited. If you are not the intended recipient, please
> contact the sender by reply e-mail and destroy all copies of this original message.
>
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>
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