Re: [Histonet] Metal embedding molds-large (Diane Satterfield)

[Mayer,Toysha N via Histonet]

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" <port...@msu.edu>
To: <histonet@lists.utsouthwestern.edu>
Subject: [Histonet] NK Cell markers for Mouse in Paraffin
Message-ID: <001301d358c2$e2c000d0$a8400270$@edu>
Content-Type: text/plain;   charset="us-ascii"

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

port...@msu.edu

 



--

Message: 2
Date: Wed, 8 Nov 2017 19:42:09 +
From: Diane Satterfield <diane.satterfi...@duke.edu>
To: "'histonet@lists.utsouthwestern.edu'"
    <histonet@lists.utsouthwestern.edu>
Subject: [Histonet] Metal embedding molds-large
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<blupr05mb1924e6e4a95339cadeddeec491...@blupr05mb1924.namprd05.prod.outlook.com>

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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.satterfi...@duke.edu<mailto:diane.satterfi...@duke.edu>
office  919-684-4642
pager  919-970-7328
fax  919-684-4975

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Medicine policies on confidentiality and/or with the approval of the sender.




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Message: 3
Date: Wed, 8 Nov 2017 12:23:58 -0800
From: Jay Lundgren <jaylundg...@gmail.com>
To: Diane Satterfield <diane.satterfi...@duke.edu>
Cc: "histonet@lists.utsouthwestern.edu"
<histonet@lists.utsouthwestern.edu>
Subject: Re: [Histonet] Metal embedding molds-large
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Content-Type: text/plain; charset="UTF-8"

mold release

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Re: [Histonet] Metal embedding molds-large

[Caroline Miller via Histonet]

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@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.satterfi...@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.
>>>
>>>
>>> ___
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>>>
>>
>
> ___
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-- 
Caroline Miller (mills)
Director of Histology
3Scan.com
415 2187297
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Re: [Histonet] Metal embedding molds-large

[Bryan Llewellyn via Histonet]

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.satterfi...@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.


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Re: [Histonet] Metal embedding molds-large

[Bryan Llewellyn via Histonet]

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.satterfi...@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.


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Re: [Histonet] Metal embedding molds-large

[Jay Lundgren via Histonet]

mold release


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On Wed, Nov 8, 2017 at 11:42 AM, Diane Satterfield via Histonet <
histonet@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.satterfi...@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.
>
>
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[Histonet] Metal embedding molds-large

[Diane Satterfield via Histonet]

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.satterfi...@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.


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