[Histonet] Metal embedding molds-large (Diane Satterfield)
Mayer,Toysha N
TNMayer at mdanderson.org
Thu Nov 9 15:35:34 CST 2017
You could boil molds in soapy water, cool, remove paraffin, rinse and dry.
You could place them in the processor on the clean cycle.
You could soak them in xylene, rinse in 100% etoh.
Milestone has a paraffin removal instrument that can be used as well. We just got one for our student lab and it is wonderful.
Toysha Mayer
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Today's Topics:
1. NK Cell markers for Mouse in Paraffin (Amy Porter)
2. Metal embedding molds-large (Diane Satterfield)
3. Re: Metal embedding molds-large (Jay Lundgren)
4. Re: Metal embedding molds-large (Bryan Llewellyn)
5. Re: Metal embedding molds-large (Bryan Llewellyn)
6. Re: Metal embedding molds-large (Caroline Miller)
7. Re: Elastic Stain (Caroline Miller)
8. Chrome Alum slides picking up eosin non specifically?
(Kathleen S Cormier)
----------------------------------------------------------------------
Message: 1
Date: Wed, 8 Nov 2017 13:53:37 -0500
From: "Amy Porter" <portera at msu.edu>
To: <histonet at lists.utsouthwestern.edu>
Subject: [Histonet] NK Cell markers for Mouse in Paraffin
Message-ID: <001301d358c2$e2c000d0$a8400270$@edu>
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Hi all - anyone out there have an antibody that they like for NK Cells in a mouse model - I am trying to work with a mouse monoclonal (MOM) with polymer technology. Looking for assistance from labs that have established protocols. We have tried enzymatic and heat retrieval ..not getting good results. Any comments would be appreciated - willing to change antibody vendors
currently using clone PK136. Thanks - Amy
Amy S. Porter, HT (ASCP)
Michigan State University - Department of Physiology
Investigative HistoPathology Lab - Supervisor
Research Core Support Facility
567 Wilson Road - Room 2201
East Lansing, MI 48824-6458
517-884-5026
portera at msu.edu
------------------------------
Message: 2
Date: Wed, 8 Nov 2017 19:42:09 +0000
From: Diane Satterfield <diane.satterfield at duke.edu>
To: "'histonet at lists.utsouthwestern.edu'"
<histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Metal embedding molds-large
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We are using large metal molds to embed mouse brains. We are having a hard time getting to block out of the molds, the paraffin blocks are sticking. Sometimes they are coming out cracked. Sometimes the cassette comes off the paraffin block. Any idea why this is happening? Any advice on how to fix this problem?
Diane L. Satterfield, BS
Manager Brain Tumor BioRepository
Research Program Leader
Duke University Medical Center
Brain Tumor Center Biorepository and Database
diane.satterfield at duke.edu<mailto:diane.satterfield at duke.edu>
office 919-684-4642
pager 919-970-7328
fax 919-684-4975
CONFIDENTIALITY NOTICE: The information contained in this electronic mail is sensitive, protected information intended only for the addressee(s). Any other person, including anyone who believes he/she might have received it due to an addressing error, is requested to notify the sender immediately by return electronic mail, and to delete it without further reading or retention. The information is not to be forwarded to or shared unless in compliance with Duke Medicine policies on confidentiality and/or with the approval of the sender.
------------------------------
Message: 3
Date: Wed, 8 Nov 2017 12:23:58 -0800
From: Jay Lundgren <jaylundgren at gmail.com>
To: Diane Satterfield <diane.satterfield at duke.edu>
Cc: "histonet at lists.utsouthwestern.edu"
<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Metal embedding molds-large
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On Wed, Nov 8, 2017 at 11:42 AM, Diane Satterfield via Histonet < histonet at lists.utsouthwestern.edu> wrote:
> We are using large metal molds to embed mouse brains. We are having a
> hard time getting to block out of the molds, the paraffin blocks are
> sticking. Sometimes they are coming out cracked. Sometimes the
> cassette comes off the paraffin block. Any idea why this is
> happening? Any advice on how to fix this problem?
>
>
> Diane L. Satterfield, BS
> Manager Brain Tumor BioRepository
> Research Program Leader
> Duke University Medical Center
> Brain Tumor Center Biorepository and Database
>
> diane.satterfield at duke.edu<mailto:diane.satterfield at duke.edu>
> office 919-684-4642
> pager 919-970-7328
> fax 919-684-4975
>
> CONFIDENTIALITY NOTICE: The information contained in this electronic
> mail is sensitive, protected information intended only for the addressee(s).
> Any other person, including anyone who believes he/she might have
> received it due to an addressing error, is requested to notify the
> sender immediately by return electronic mail, and to delete it without
> further reading or retention. The information is not to be forwarded
> to or shared unless in compliance with Duke Medicine policies on
> confidentiality and/or with the approval of the sender.
>
>
> _______________________________________________
> Histonet mailing list
> Histonet at lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
------------------------------
Message: 4
Date: Wed, 8 Nov 2017 12:24:08 -0800
From: Bryan Llewellyn <llewllew at shaw.ca>
To: Diane Satterfield <diane.satterfield at duke.edu>, Histonet
<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Metal embedding molds-large
Message-ID: <11e6f002-2f10-63ac-8fa4-691bdbf847f7 at shaw.ca>
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This used to be a common problem years ago. It is due to crud buildup on the metal. Boil them with TCP for half an hour, then thoroughly wash them in cold water. Coat them with a VERY light smear of glycerol before you use them, preferably each time. That should help.
Bryan Llewellyn.
Diane Satterfield via Histonet wrote:
> We are using large metal molds to embed mouse brains. We are having a hard time getting to block out of the molds, the paraffin blocks are sticking. Sometimes they are coming out cracked. Sometimes the cassette comes off the paraffin block. Any idea why this is happening? Any advice on how to fix this problem?
>
>
> Diane L. Satterfield, BS
> Manager Brain Tumor BioRepository
> Research Program Leader
> Duke University Medical Center
> Brain Tumor Center Biorepository and Database
>
> diane.satterfield at duke.edu<mailto:diane.satterfield at duke.edu>
> office 919-684-4642
> pager 919-970-7328
> fax 919-684-4975
>
> CONFIDENTIALITY NOTICE: The information contained in this electronic mail is sensitive, protected information intended only for the addressee(s). Any other person, including anyone who believes he/she might have received it due to an addressing error, is requested to notify the sender immediately by return electronic mail, and to delete it without further reading or retention. The information is not to be forwarded to or shared unless in compliance with Duke Medicine policies on confidentiality and/or with the approval of the sender.
>
>
> _______________________________________________
> Histonet mailing list
> Histonet at lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
------------------------------
Message: 5
Date: Wed, 8 Nov 2017 12:30:06 -0800
From: Bryan Llewellyn <llewllew at shaw.ca>
To: Diane Satterfield <diane.satterfield at duke.edu>, Histonet
<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Metal embedding molds-large
Message-ID: <e30eeb29-90fe-557a-609f-c47891e13c40 at shaw.ca>
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Sorry!
That should be TSP - trisodium phosphate - not TCP, which might make it worse.
Bryan
Bryan Llewellyn wrote:
> This used to be a common problem years ago. It is due to crud buildup
> on the metal. Boil them with TCP for half an hour, then thoroughly
> wash them in cold water. Coat them with a VERY light smear of glycerol
> before you use them, preferably each time. That should help.
>
> Bryan Llewellyn.
>
> Diane Satterfield via Histonet wrote:
>> We are using large metal molds to embed mouse brains. We are having
>> a hard time getting to block out of the molds, the paraffin blocks
>> are sticking. Sometimes they are coming out cracked. Sometimes the
>> cassette comes off the paraffin block. Any idea why this is
>> happening? Any advice on how to fix this problem?
>>
>>
>> Diane L. Satterfield, BS
>> Manager Brain Tumor BioRepository
>> Research Program Leader
>> Duke University Medical Center
>> Brain Tumor Center Biorepository and Database
>>
>> diane.satterfield at duke.edu<mailto:diane.satterfield at duke.edu>
>> office 919-684-4642
>> pager 919-970-7328
>> fax 919-684-4975
>>
>> CONFIDENTIALITY NOTICE: The information contained in this electronic
>> mail is sensitive, protected information intended only for the
>> addressee(s). Any other person, including anyone who believes he/she
>> might have received it due to an addressing error, is requested to
>> notify the sender immediately by return electronic mail, and to
>> delete it without further reading or retention. The information is
>> not to be forwarded to or shared unless in compliance with Duke
>> Medicine policies on confidentiality and/or with the approval of the sender.
>>
>>
>> _______________________________________________
>> Histonet mailing list
>> Histonet at lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>
------------------------------
Message: 6
Date: Wed, 8 Nov 2017 13:46:39 -0800
From: Caroline Miller <mills at 3scan.com>
To: Bryan Llewellyn <llewllew at shaw.ca>
Cc: Diane Satterfield <diane.satterfield at duke.edu>, Histonet
<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Metal embedding molds-large
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<CAFTHRQOe=i5DN=-6Pnjk-7mYu+tf4=75YtCjRhuKGFfG-VbUvA at mail.gmail.com>
Content-Type: text/plain; charset="UTF-8"
hey, we do all our embedding in those molds, and here is what I suggest:
1 - Make sure to have enough wax in the back of the molds, all the way until it is on the lip of the cassette - you may need to refill a bunch of times because the wax drains out (why regular sakura cassettes do not fit in these molds, also made by sakura I really don't know). But you need to wait until it hardens enough, but not too much to leave a transition between the two fill waxes. This will prevent the cassette from coming away when you pop it out
2 - Make sure the block is nice and cold, wait 10 minutes longer than you think you have to
3 - Use a spatula or other strong item to place under the label side of the cassette and pop out of the mold, if you get any resistance then WAIT some more! (again them being cold is really important)
good luck, happy to answer any clarifying questions!
mills
On Wed, Nov 8, 2017 at 12:30 PM, Bryan Llewellyn via Histonet < histonet at lists.utsouthwestern.edu> wrote:
> Sorry!
>
> That should be TSP - trisodium phosphate - not TCP, which might make
> it worse.
>
> Bryan
>
>
>
> Bryan Llewellyn wrote:
>
>> This used to be a common problem years ago. It is due to crud buildup
>> on the metal. Boil them with TCP for half an hour, then thoroughly
>> wash them in cold water. Coat them with a VERY light smear of
>> glycerol before you use them, preferably each time. That should help.
>>
>> Bryan Llewellyn.
>>
>> Diane Satterfield via Histonet wrote:
>>
>>> We are using large metal molds to embed mouse brains. We are having
>>> a hard time getting to block out of the molds, the paraffin blocks
>>> are sticking. Sometimes they are coming out cracked. Sometimes the
>>> cassette comes off the paraffin block. Any idea why this is
>>> happening? Any advice on how to fix this problem?
>>>
>>>
>>> Diane L. Satterfield, BS
>>> Manager Brain Tumor BioRepository
>>> Research Program Leader
>>> Duke University Medical Center
>>> Brain Tumor Center Biorepository and Database
>>>
>>> diane.satterfield at duke.edu<mailto:diane.satterfield at duke.edu>
>>> office 919-684-4642
>>> pager 919-970-7328
>>> fax 919-684-4975
>>>
>>> CONFIDENTIALITY NOTICE: The information contained in this
>>> electronic mail is sensitive, protected information intended only
>>> for the addressee(s). Any other person, including anyone who
>>> believes he/she might have received it due to an addressing error,
>>> is requested to notify the sender immediately by return electronic
>>> mail, and to delete it without further reading or retention. The
>>> information is not to be forwarded to or shared unless in compliance
>>> with Duke Medicine policies on confidentiality and/or with the approval of the sender.
>>>
>>>
>>> _______________________________________________
>>> Histonet mailing list
>>> Histonet at lists.utsouthwestern.edu
>>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>>
>>>
>>
>
> _______________________________________________
> Histonet mailing list
> Histonet at lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
--
Caroline Miller (mills)
Director of Histology
3Scan.com
415 2187297
------------------------------
Message: 7
Date: Wed, 8 Nov 2017 13:48:13 -0800
From: Caroline Miller <mills at 3scan.com>
To: Terri Braud <tbraud at holyredeemer.com>
Cc: "histonet at lists.utsouthwestern.edu"
<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Elastic Stain
Message-ID:
<CAFTHRQP5OYoOf0QqtMOmwGbrsrSr5oWfRgh7ZPJtKf-3O0QtsQ at mail.gmail.com>
Content-Type: text/plain; charset="UTF-8"
In the UK clinical lab I started in we used Miller's elastin, which I think
was a resourcinol formulation (digging from 20 years ago in my brain). I
tried to find that in the US but never did. I loved that stain, worked well
every time! We would counterstain with a van-Giesen, looked stunning! the
elastin came out very dark blue / black.
mills
On Wed, Nov 8, 2017 at 6:31 AM, Terri Braud via Histonet <
histonet at lists.utsouthwestern.edu> wrote:
> We love the Aldehyde Fuchsin with a fast-green counterstain. Once the
> initial working stain is made up, it is a fast and easy stain to perform,
> not to mention just pretty - purple fibers against a green background.
>
> Terri L. Braud, HT(ASCP)
> Anatomic Pathology Supervisor
> Laboratory
> Holy Redeemer Hospital
> 1648 Huntingdon Pike
> Meadowbrook, PA 19046
> ph: 215-938-3689
> fax: 215-938-3874
> Care, Comfort, and Heal
> *************************
>
>
> _______________________________________________
> Histonet mailing list
> Histonet at lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
--
Caroline Miller (mills)
Director of Histology
3Scan.com
415 2187297
------------------------------
Message: 8
Date: Thu, 9 Nov 2017 12:23:33 +0000
From: Kathleen S Cormier <cormier at mit.edu>
To: "histonet at lists.utsouthwestern.edu"
<histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Chrome Alum slides picking up eosin non
specifically?
Message-ID: <68218F88-E8FA-4D49-82D6-ECAB218B507C at mit.edu>
Content-Type: text/plain; charset="us-ascii"
Hello Everyone,
Was wondering, I am making up some chrome alum slides, and they seem to pick up quite the bit of eosin non specifically. Is this typical? If not, what should I do differently? Thanks!
Kathy
Kathy Cormier
cormier at mit.edu<mailto:cormier at mit.edu>
------------------------------
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