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 -----Original Message----- From: histonet-requ...@lists.utsouthwestern.edu [mailto:histonet-requ...@lists.utsouthwestern.edu] Sent: Thursday, November 09, 2017 12:00 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 168, Issue 8 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-requ...@lists.utsouthwestern.edu You can reach the person managing the list at histonet-ow...@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." 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 +0000 From: Diane Satterfield <diane.satterfi...@duke.edu> To: "'histonet@lists.utsouthwestern.edu'" <histonet@lists.utsouthwestern.edu> Subject: [Histonet] Metal embedding molds-large Message-ID: <blupr05mb1924e6e4a95339cadeddeec491...@blupr05mb1924.namprd05.prod.outlook.com> Content-Type: text/plain; charset="us-ascii" 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 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 <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 Message-ID: <cancznuzk3wrp+ut9sdhrjfybx_b6_w_zpbzmxwbuav6t2na...@mail.gmail.com> Content-Type: text/plain; charset="UTF-8" mold release <http://www.avg.com/email-signature?utm_medium=email&utm_source=link&utm_campaign=sig-email&utm_content=webmail> Virus-free. www.avg.com <http://www.avg.com/email-signature?utm_medium=email&utm_source=link&utm_campaign=sig-email&utm_content=webmail> <#DAB4FAD8-2DD7-40BB-A1B8-4E2AA1F9FDF2> 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<mailto: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. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 4 Date: Wed, 8 Nov 2017 12:24:08 -0800 From: Bryan Llewellyn <llewl...@shaw.ca> To: Diane Satterfield <diane.satterfi...@duke.edu>, Histonet <histonet@lists.utsouthwestern.edu> Subject: Re: [Histonet] Metal embedding molds-large Message-ID: <11e6f002-2f10-63ac-8fa4-691bdbf84...@shaw.ca> Content-Type: text/plain; charset=UTF-8; format=flowed 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<mailto: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. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 5 Date: Wed, 8 Nov 2017 12:30:06 -0800 From: Bryan Llewellyn <llewl...@shaw.ca> To: Diane Satterfield <diane.satterfi...@duke.edu>, Histonet <histonet@lists.utsouthwestern.edu> Subject: Re: [Histonet] Metal embedding molds-large Message-ID: <e30eeb29-90fe-557a-609f-c47891e13...@shaw.ca> Content-Type: text/plain; charset=UTF-8; format=flowed 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<mailto: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. >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > ------------------------------ Message: 6 Date: Wed, 8 Nov 2017 13:46:39 -0800 From: Caroline Miller <mi...@3scan.com> To: Bryan Llewellyn <llewl...@shaw.ca> Cc: Diane Satterfield <diane.satterfi...@duke.edu>, Histonet <histonet@lists.utsouthwestern.edu> Subject: Re: [Histonet] Metal embedding molds-large Message-ID: <CAFTHRQOe=i5DN=-6Pnjk-7mYu+tf4=75ytcjrhukgffg-vb...@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@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<mailto: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. >>> >>> >>> _______________________________________________ >>> Histonet mailing list >>> Histonet@lists.utsouthwestern.edu >>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >>> >>> >> > > _______________________________________________ > Histonet mailing list > Histonet@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 <mi...@3scan.com> To: Terri Braud <tbr...@holyredeemer.com> Cc: "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> Subject: Re: [Histonet] Elastic Stain Message-ID: <cafthrqp5oyoof0qqtmomwgbrsrsr5owfrgh7zpjtkf-3o0q...@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@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@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 <corm...@mit.edu> To: "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> Subject: [Histonet] Chrome Alum slides picking up eosin non specifically? Message-ID: <68218f88-e8fa-4d49-82d6-ecab218b5...@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 corm...@mit.edu<mailto:corm...@mit.edu> ------------------------------ Subject: Digest Footer _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ End of Histonet Digest, Vol 168, Issue 8 **************************************** The information contained in this e-mail message may be privileged, confidential, and/or protected from disclosure. This e-mail message may contain protected health information (PHI); dissemination of PHI should comply with applicable federal and state laws. If you are not the intended recipient, or an authorized representative of the intended recipient, any further review, disclosure, use, dissemination, distribution, or copying of this message or any attachment (or the information contained therein) is strictly prohibited. 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