[Histonet] slide scanner
Hi, I am looking to buy a slide scanner for highthrouput project. Could you give me your opinion if you are using one of the next 3: Mirax Scan Zeiss, Nanozoomer Hamamatsu and XT Aperio? Best regards Jeanne Jeanne Estabel, PhD MGP Histology Operations Manager Wellcome Trust/Sanger Institute UK -- The Wellcome Trust Sanger Institute is operated by Genome Research Limited, a charity registered in England with number 1021457 and a compa ny registered in England with number 2742969, whose registered office is 2 15 Euston Road, London, NW1 2BE. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] HRP-labeled primary antibodies
Hi All, Has anyone had experience doing IHC on FFPE tissues with HRP-labeled primary antibodies? I was wondering what the best way to detect them would be. I assume that going strait to DAB would not work, since no amplification is there. I was thinking of using a biotinyl tyramide step to amplify the signal. Also, do you think the final antibody concentration would need to be higher than with traditional, unlabeled primaries? Thanks, Kim Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Gross photography/macrophotography
I have been very happy with the Olympus C-7000. It will focus at 3 1/4 inches (8 cm) in its macro mode. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Reuel Cornelia Sent: Wednesday, May 20, 2009 4:53 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Gross photography/macrophotography Hello histonetters, I am looking for a good digital camera for gross photography. Any recommendations that works with your lab will be beneficial. Thank you. Happy memorial day!!! Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 ___ 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
[Histonet] Bone!
Friends, Can someone recommend to me how to store bones (juvenile rat femur) that I have fixed and decalcified, but am not yet ready to cut (they're for vibratome, so not embedded). Thanks! Redward. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] bone fixation and calcification
Dear Histonetters, I am new in fixation and processing bones. I need your full protocol with details how to fix and process mice bone to visualize the tumor metastases? Thanks in advance, Naira ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] HRP-labeled primary antibodies
If there is enough of the antigen in the tissue or sample you can detect without amplification. HRP/DAB is an enzymatic reaction so it is also is amplification. I would try it the straight up way before diving into more complex protocols. Alternatively another option would be that you could try to come in with a secondary antibody (either HRP labeled itself or biotinylated). If your secondary is a polyclonal - which most are - it should still be able to detect the primary even with the HRP attached. Worth trying anyway. -Original Message- From: Kim Merriam kmerriam2...@yahoo.com Date: Thu, 21 May 2009 05:41:44 To: Histonethistonet@lists.utsouthwestern.edu; ih...@googlegroups.com Subject: [Histonet] HRP-labeled primary antibodies Hi All, Has anyone had experience doing IHC on FFPE tissues with HRP-labeled primary antibodies? I was wondering what the best way to detect them would be. I assume that going strait to DAB would not work, since no amplification is there. I was thinking of using a biotinyl tyramide step to amplify the signal. Also, do you think the final antibody concentration would need to be higher than with traditional, unlabeled primaries? Thanks, Kim Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA ___ 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
Re: [Histonet] HRP-labeled primary antibodies
I got some great ideas. Thanks everyone! Kim Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA From: anh2...@med.cornell.edu anh2...@med.cornell.edu To: Kim Merriam kmerriam2...@yahoo.com; Histonet histonet@lists.utsouthwestern.edu; ih...@googlegroups.com Sent: Thursday, May 21, 2009 10:48:15 AM Subject: Re: [Histonet] HRP-labeled primary antibodies If there is enough of the antigen in the tissue or sample you can detect without amplification. HRP/DAB is an enzymatic reaction so it is also is amplification. I would try it the straight up way before diving into more complex protocols. Alternatively another option would be that you could try to come in with a secondary antibody (either HRP labeled itself or biotinylated). If your secondary is a polyclonal - which most are - it should still be able to detect the primary even with the HRP attached. Worth trying anyway. -Original Message- From: Kim Merriam kmerriam2...@yahoo.com Date: Thu, 21 May 2009 05:41:44 To: Histonethistonet@lists.utsouthwestern.edu; ih...@googlegroups.com Subject: [Histonet] HRP-labeled primary antibodies Hi All, Has anyone had experience doing IHC on FFPE tissues with HRP-labeled primary antibodies? I was wondering what the best way to detect them would be. I assume that going strait to DAB would not work, since no amplification is there. I was thinking of using a biotinyl tyramide step to amplify the signal. Also, do you think the final antibody concentration would need to be higher than with traditional, unlabeled primaries? Thanks, Kim Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA ___ 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
[Histonet] bone tissue
Can somebody help me with my bone tissue to remain intact during antigen retrieval for IHC staining. I have used subbed slides but It did not help me. Is there more better adhesive. Does Haupts Gelatin works better? I know this is a hundred year old question and hopefully this problem have been resolved since my undated knowledge could remember. I would like to thank everybody for responding to my last e-mail on gross photography. It gives me more idea what to purchase. Thanks again histonetters and I really appreciate your help. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Bone!
Thanks Kemlo... I thought of that, but was worried about over-fixation,- what do you think? Redward. -- In 10% neutral buffered formalin? Kemlo Rogerson e-mail kemloroger...@nhs.net if not at work. DD 01934 647057 or extension 3311 Mob 07749 754194; Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah Lehrer This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Robert Edward Pogue Sent: 21 May 2009 14:47 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone! Friends, Can someone recommend to me how to store bones (juvenile rat femur) that I have fixed and decalcified, but am not yet ready to cut (they're for vibratome, so not embedded). Thanks! Redward. ___ 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
RE: [Histonet] Bone saw
IMEB also has a Bone Band Saw. http://www.imebinc.com/Item/BBS-82203.htm Jack Date: Thu, 21 May 2009 09:05:50 -0700 From: cb...@memorialcare.org To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone saw Good Morning, I was wondering if anyone can help me find a really good saw for bones (femoral/humeral heads mainly). We currently have a MarMed bone saw that works great for knees and such but it's just not strong enough for the femurs. Thank you, Christine Bark HT(ASCP) CM Lead Histotech, Pathology Saddleback Memorial Medical Center 949-452-3548 cb...@memorialcare.org __ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and 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 the original message. ___ 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
[Histonet] cloudy formalin in tissue processor
Has anyone had a problem with their formalin getting cloudy in their tissue processor? We have a VIP tissue processor and just recently started having problems with cloudy formalin. Any suggestions would be appreciated. I checked the archives and couldn't find any articles on this particular subject. Thanks. CONFIDENTIALITY NOTICE: This electronic message, including any attachments, is for the sole use of the intended recipients and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by electronic mail and destroy all copies of the original message. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] bone tissue
Haupts Gelatin works really well at a 1:1 ratio (Concentrate:50% EtOH). The only limitation is background staining from say a hematoxylin counterstain after the IHC. You can purchase the concentrate from Fisher (Haupts Fixative #785-71). Jack Date: Thu, 21 May 2009 10:12:23 -0500 From: reuel.corne...@tsrh.org To: histonet@lists.utsouthwestern.edu Subject: [Histonet] bone tissue Can somebody help me with my bone tissue to remain intact during antigen retrieval for IHC staining. I have used subbed slides but It did not help me. Is there more better adhesive. Does Haupts Gelatin works better? I know this is a hundred year old question and hopefully this problem have been resolved since my undated knowledge could remember. I would like to thank everybody for responding to my last e-mail on gross photography. It gives me more idea what to purchase. Thanks again histonetters and I really appreciate your help. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 ___ 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
RE: [Histonet] Bone!
I do not routinely perform decalcified bone preparations as I predominantly utilize undemineralized resin (MMA) applications, so maybe someone else can better respond to this question. However, I would say that you could store your bone in 70% EtOH until you are ready to process, but the length of time in solution storage may cause you problems down the road since you have already decalcified. I think that a prolonged storage time might make your bone hard againif that makes any senseand cause you a fit later when you go to the microtome. Typically, I store undemineralized bones at either 10% NBF or 70% EtOH depending upon what I care to see at the microscope. Jack Date: Thu, 21 May 2009 10:46:59 -0300 From: redw...@ucb.br To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone! Friends, Can someone recommend to me how to store bones (juvenile rat femur) that I have fixed and decalcified, but am not yet ready to cut (they're for vibratome, so not embedded). Thanks! Redward. ___ 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
RE: [IHCRG] Re: [Histonet] HRP-labeled primary antibodies
I agree with Kim, if I was worried about a weak signal using abs directly
conjugated to hrp I would just ignore the fact that they have the hrp and
use a detection such as labeled polymer matched to the species of the
primary ab just like I would without the direct hrp conjugation, there
should still be enough sites left on the primary antibody for a secondary
detection reagent to attach to the species ab even with some being taken up
by the hrp.
Patsy
Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 215
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pru...@ihctech.net
web site www.ihctech.net
This email is confidential and intended solely for the use of the Person(s)
('the intended recipient') to whom it was addressed. Any views or opinions
presented are solely those of the author. It may contain information that is
privileged confidential within the meaning of applicable law. Accordingly
any dissemination, distribution, copying, or other use of this message, or
any of its contents, by any person other than the intended recipient may
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From: ih...@googlegroups.com [mailto:ih...@googlegroups.com] On Behalf Of
Kim Merriam
Sent: Thursday, May 21, 2009 8:52 AM
To: Histonet; ih...@googlegroups.com
Subject: [IHCRG] Re: [Histonet] HRP-labeled primary antibodies
I got some great ideas. Thanks everyone!
Kim
Kim Merriam, MA, HT(ASCP)QIHC
Cambridge, MA
_
From: anh2...@med.cornell.edu anh2...@med.cornell.edu
To: Kim Merriam kmerriam2...@yahoo.com; Histonet
histonet@lists.utsouthwestern.edu; ih...@googlegroups.com
Sent: Thursday, May 21, 2009 10:48:15 AM
Subject: Re: [Histonet] HRP-labeled primary antibodies
If there is enough of the antigen in the tissue or sample you can detect
without amplification. HRP/DAB is an enzymatic reaction so it is also is
amplification. I would try it the straight up way before diving into more
complex protocols.
Alternatively another option would be that you could try to come in with a
secondary antibody (either HRP labeled itself or biotinylated). If your
secondary is a polyclonal - which most are - it should still be able to
detect the primary even with the HRP attached. Worth trying anyway.
-Original Message-
From: Kim Merriam kmerriam2...@yahoo.com
Date: Thu, 21 May 2009 05:41:44
To: Histonethistonet@lists.utsouthwestern.edu; ih...@googlegroups.com
Subject: [Histonet] HRP-labeled primary antibodies
Hi All,
Has anyone had experience doing IHC on FFPE tissues with HRP-labeled primary
antibodies? I was wondering what the best way to detect them would be. I
assume that going strait to DAB would not work, since no amplification is
there. I was thinking of using a biotinyl tyramide step to amplify the
signal.
Also, do you think the final antibody concentration would need to be higher
than with traditional, unlabeled primaries?
Thanks,
Kim
Kim Merriam, MA, HT(ASCP)QIHC
Cambridge, MA
___
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[Histonet] fume hood regulations
Fellow Histonetters, Does anyone have knowledge of guidelines or regulations on whether a fume hood must remain on 24/7? In a cost cutting measure a facility is considering turning off the fume hoods from 7pm to 5am. I have expressed my concern with the idea because of the huge health and safety issue for those who work after hours. This facility could be setting themselves up for a huge liability law suit. I would appreciate written guidelines to strengthen my opposition to this cost cutting measure. Thank you, Tina ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] cloudy formalin in tissue processor
Is your formalin picking up water from the air in the room? Check the humidity level in your room. Lynn Burton Lab Assoc. I Animal Disease Lab Galesburg, Il 61401 From: histonet-boun...@lists.utsouthwestern.edu on behalf of Christine I. Braaten Sent: Thu 5/21/2009 11:41 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cloudy formalin in tissue processor Has anyone had a problem with their formalin getting cloudy in their tissue processor? We have a VIP tissue processor and just recently started having problems with cloudy formalin. Any suggestions would be appreciated. I checked the archives and couldn't find any articles on this particular subject. Thanks. CONFIDENTIALITY NOTICE: This electronic message, including any attachments, is for the sole use of the intended recipients and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by electronic mail and destroy all copies of the original message. ___ 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
Re: [Histonet] fume hood regulations
Hi, At the University of Washington we have been told to lower the sash all the way when the hood is not in use to save energy and money. I believe there have been several Universities that have done some cost analysis of this and have show large savings in money (one study stated $1500 per year per fume hood in savings and a reduction in 10,600 lbs of CO2 emissions). If you want more information just do a search for shut the sash. So, if it is not feasible to shut off the hood you can save energy and money just by closing the sash. Have a great weekend. Yak-Nam On 5/21/09 10:10 AM, Montina Van Meter montina.vanme...@pbrc.edu wrote: Fellow Histonetters, Does anyone have knowledge of guidelines or regulations on whether a fume hood must remain on 24/7? In a cost cutting measure a facility is considering turning off the fume hoods from 7pm to 5am. I have expressed my concern with the idea because of the huge health and safety issue for those who work after hours. This facility could be setting themselves up for a huge liability law suit. I would appreciate written guidelines to strengthen my opposition to this cost cutting measure. Thank you, Tina ___ 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
[Histonet] IHC question- freezing aliquots
Hello all! I have an antibody that has a 2 month expiration date. It is recommended that we aliquot, and freeze (-80) the antibody- per the vendor. How do you interpret: The CAP reg pertaining to expired antibody use: ANP .22432 ??? Are all immunohistochemical reagents used within their indicated dates? How are other institutions managing their antibodies that require freezing.--- do you aliquot,freeze, and give the reagent an extended expiration after it has been diluted? Thanks in advance!!! I have read the past posts on this in the archives, but am wondering if there is a clear answer to antibody use and freezing. Theresa R Dobersztyn HT (ASCP) Electron Microscopy/Histology Labs Senior Technologist Department of Pathology Akron Children's Hospital 1 Perkins Square Akron, Ohio 44308 330-543-8279 Akron Children's Hospital - Proud winner of the NorthCoast 99 Best Workplace award! * This electronic mail transmission, including any attached files, may contain confidential and/or privileged information for the sole use of the intended recipient(s). It is not intended for transmission to, or receipt by, any unauthorized parties. Any review, use, distribution, dissemination, downloading, copying or disclosure by others is strictly prohibited. If you are not the intended recipient (or authorized to receive information for the intended recipient), you are hereby notified that you have received this transmission in error. If you have received this transmission in error, please delete it, as well as any copies, from your system without copying it, and notify the sender by reply e-mail. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Dry ice - transportation
Are there any restrictions on the transportation of dry ice (and human tissue) in private vehicles? Our PAs cover a surgical center off-site and we need to transport frozen tissue back to the hospital for biobanking purposes. They drive back and forth in their own vehicles. Thanks. Richard Richard W. Cartun, Ph.D. Director, Histology Immunopathology Director, Biospecimens Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IHC question- freezing aliquots
In a similar situation we have gotten documentation (e-mail) from the vendor stating the recommendation to freeze aliquots and to what degree that extends the shelf life. We haven't been queried on these particular antibodies by an inspector but our supervisor feels that this satisfies the CAP reg. Linda A. Sebree University of Wisconsin Hospital Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of tdobersz...@chmca.org Sent: Thursday, May 21, 2009 1:19 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC question- freezing aliquots Hello all! I have an antibody that has a 2 month expiration date. It is recommended that we aliquot, and freeze (-80) the antibody- per the vendor. How do you interpret: The CAP reg pertaining to expired antibody use: ANP .22432 ??? Are all immunohistochemical reagents used within their indicated dates? How are other institutions managing their antibodies that require freezing.--- do you aliquot,freeze, and give the reagent an extended expiration after it has been diluted? Thanks in advance!!! I have read the past posts on this in the archives, but am wondering if there is a clear answer to antibody use and freezing. Theresa R Dobersztyn HT (ASCP) Electron Microscopy/Histology Labs Senior Technologist Department of Pathology Akron Children's Hospital 1 Perkins Square Akron, Ohio 44308 330-543-8279 Akron Children's Hospital - Proud winner of the NorthCoast 99 Best Workplace award! * This electronic mail transmission, including any attached files, may contain confidential and/or privileged information for the sole use of the intended recipient(s). It is not intended for transmission to, or receipt by, any unauthorized parties. Any review, use, distribution, dissemination, downloading, copying or disclosure by others is strictly prohibited. If you are not the intended recipient (or authorized to receive information for the intended recipient), you are hereby notified that you have received this transmission in error. If you have received this transmission in error, please delete it, as well as any copies, from your system without copying it, and notify the sender by reply e-mail. ___ 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
[Histonet] Plant/insect histology
If anyone out in histonet land is doing above, could you please contact me by email directly? Thanks Luis /PRE html body br / This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain information that is proprietary, confidential, and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure, or distribution is prohibited. If you have received this email in error please notify the sender by return email and delete the original message. Please note, the recipient should check this email and any attachments for the presence of viruses. The organization accepts no liability for any damage caused by any virus transmitted by this email.br / = /body /html PRE ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Dry ice - transportation
Richard: There may be local regulations that could vary. Here you can buy dry ice in the grocery store (if you bring an appropriate container). The safety warnings associated with dry ice indicate (as you would expect) - transport in unsealed insulated container (Styrofoam chest), transport in the trunk of your car not the passenger compartment and crack a window, avoid direct contact with skin eyes or ingestion to avoid burns, etc. If your PA's are traveling long distances using a minivan SUV or similar single compartment vehicle, the main problem would be accumulation of CO2 gas that would ultimately induce drowsiness and in worst case scenario asphyxia. Joe -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Richard Cartun Sent: Thursday, May 21, 2009 1:33 PM To: Histonet Subject: [Histonet] Dry ice - transportation Are there any restrictions on the transportation of dry ice (and human tissue) in private vehicles? Our PAs cover a surgical center off-site and we need to transport frozen tissue back to the hospital for biobanking purposes. They drive back and forth in their own vehicles. Thanks. Richard Richard W. Cartun, Ph.D. Director, Histology Immunopathology Director, Biospecimens Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ___ 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
[Histonet] Adhesive for immunos
We have used 5% Tite-bond glue( from the Hardware store) on slides with great success. Just take a Kimwipe and wipe it evenly over the slide and it holds the tissue very well. Annette Featherstone, Supervisor of Pathology and Anatomical Sciences The University of Buffalo Phone:716-829-3108 Fax: 716-829-2911 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] cloudy formalin in tissue processor
If you use NBF it will get cloudy when it mixes (in some step) with alcohol. The NBF has salts that are not soluble in alcohol and will produce cloudiness. You are getting your NBF mixed with alcohol. René J. --- On Thu, 5/21/09, Burton, Lynn lynn.bur...@illinois.gov wrote: From: Burton, Lynn lynn.bur...@illinois.gov Subject: RE: [Histonet] cloudy formalin in tissue processor To: Christine I. Braaten cbraa...@cheshire-med.com, histonet@lists.utsouthwestern.edu Date: Thursday, May 21, 2009, 1:23 PM Is your formalin picking up water from the air in the room? Check the humidity level in your room. Lynn Burton Lab Assoc. I Animal Disease Lab Galesburg, Il 61401 From: histonet-boun...@lists.utsouthwestern.edu on behalf of Christine I. Braaten Sent: Thu 5/21/2009 11:41 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cloudy formalin in tissue processor Has anyone had a problem with their formalin getting cloudy in their tissue processor? We have a VIP tissue processor and just recently started having problems with cloudy formalin. Any suggestions would be appreciated. I checked the archives and couldn't find any articles on this particular subject. Thanks. CONFIDENTIALITY NOTICE: This electronic message, including any attachments, is for the sole use of the intended recipients and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by electronic mail and destroy all copies of the original message. ___ 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] cloudy formalin in tissue processor
This can result from not drying the retort or tissue tank after the clean cycle which will leave alcohol in the bottom. Dry the tank beofre starting the next run. Ocassionally I have even seen some xylene end up in a mixture in the tissue tank if it is not dried well. Both of these will turn your NBF cloudy. When you get a processor they don't always tell you to dry it to prevent carry over. Pam Marcum UPENN Vet Sch New Bolton Center - Original Message - From: Rene J Buesa rjbu...@yahoo.com To: Christine I. Braaten cbraa...@cheshire-med.com, histonet@lists.utsouthwestern.edu, LynnBurton lynn.bur...@illinois.gov Sent: Thursday, May 21, 2009 3:49:06 PM GMT -05:00 US/Canada Eastern Subject: RE: [Histonet] cloudy formalin in tissue processor If you use NBF it will get cloudy when it mixes (in some step) with alcohol. The NBF has salts that are not soluble in alcohol and will produce cloudiness. You are getting your NBF mixed with alcohol. René J. --- On Thu, 5/21/09, Burton, Lynn lynn.bur...@illinois.gov wrote: From: Burton, Lynn lynn.bur...@illinois.gov Subject: RE: [Histonet] cloudy formalin in tissue processor To: Christine I. Braaten cbraa...@cheshire-med.com, histonet@lists.utsouthwestern.edu Date: Thursday, May 21, 2009, 1:23 PM Is your formalin picking up water from the air in the room? Check the humidity level in your room. Lynn Burton Lab Assoc. I Animal Disease Lab Galesburg, Il 61401 From: histonet-boun...@lists.utsouthwestern.edu on behalf of Christine I. Braaten Sent: Thu 5/21/2009 11:41 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cloudy formalin in tissue processor Has anyone had a problem with their formalin getting cloudy in their tissue processor? We have a VIP tissue processor and just recently started having problems with cloudy formalin. Any suggestions would be appreciated. I checked the archives and couldn't find any articles on this particular subject. Thanks. CONFIDENTIALITY NOTICE: This electronic message, including any attachments, is for the sole use of the intended recipients and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by electronic mail and destroy all copies of the original message. ___ 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 ___ 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
[Histonet] Chris Stephenson is out of the office.
I will be out of the office starting 05/21/2009 and will not return until 05/26/2009. Please forward all urgent messages to Eric Halpern at ext. 5852 or eric.halp...@comphealth.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Bone saw
Christine Bark asks about saws for cutting surgical pathology bone specimens like femoral heads. I posted something to Histonet about this fairly recently - here it is again. Stephen Peters, a pathologist in Hackensack NJ, describes for us a bone saw he's invented. His Web site is distinctly worth looking at, the whole thing, not just the saw. http://pathologyinnovations.com/bone_vise.htm Some other possibilities for sawing bone: Many of the small pathology services I work for have no way of sawing bone. (It's amazing how many pathologists there are who are so poorly trained that they think you decalcify a femoral head by tossing the whole thing in formalin for a month.) In that circumstance, I head for the nearest hardware store and buy a five dollar hacksaw which I leave behind at the end of the week. The Civil War vintage Satterlee amputation saw is still available, and is a serviceable hand saw that doesn't go dull quickly. (I've seen them, complete with chrome plating, at Civil War re-enactments.) This is the tool I most commonly use. One of the standard vendors offers a simple device for slabbing a femoral head, the SawBones, absurdly overpriced at $500, not something a hospital would be likely to buy for a mere pathologist. Several years ago I attended a continuing medical education program where the lecturer recommended a scroll saw, a large table saw that's about impossible to injure yourself with. At the time they cost about $100 for Made in China, otherwise $200. The disadvantage, in a cramped pathology lab, is its large footprint. You can look at these things at your local Home Depot. There's no way to cut a femoral head safely with an oscillating autopsy saw (Stryker saw), though this is probably the most common way to cut bone. I think femoral heads removed for fracture (not for osteoarthritis) really do need to be examined microscopically, because of the occasional pathologic fracture (fracture through metastatic cancer in the bone). I've seen several of these, not all with a previous cancer diagnosis. (But I see no reason to examine knee replacement material microscopically, if you know how to do the gross description properly - which admittedly most pathologists don't.) (I have no connection with any of the businesses I've mentioned.) Bob Richmond Samurai Pathologist Knoxville TN ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: [IHCRG] HRP-labeled primary antibodies
Kim, The direct IPX is the easiest method apart from a direct immunofluorescence. Very few steps: 1.Block endogenous enzyme 2.Block non-specific Ab binding 3.Put the HRP-conjugated Ab on (a an appropriate dilution, usually more concentrated than if you were using an ABC or Polymer amplification method) 4.Incubate in your usual DAB-H2O2 solution 5.Counterstain and you done. Throw in a few buffer washes in between and maybe some antigen retrieval if required. We use the direct IPX method for renal biopsies (IgG,M,A etc). See Birchall, I.W., (1996) Direct antibody method for formalin fixed, paraffin embedded renal biopsies: a comparison with the peroxidase labelled streptavidin/biotin method J. Histotechnol 19(2): 125-129. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: ih...@googlegroups.com [mailto:ih...@googlegroups.com] On Behalf Of Kim Merriam Sent: Thursday, 21 May 2009 10:42 PM To: Histonet; ih...@googlegroups.com Subject: [IHCRG] HRP-labeled primary antibodies Hi All, Has anyone had experience doing IHC on FFPE tissues with HRP-labeled primary antibodies? I was wondering what the best way to detect them would be. I assume that going strait to DAB would not work, since no amplification is there. I was thinking of using a biotinyl tyramide step to amplify the signal. Also, do you think the final antibody concentration would need to be higher than with traditional, unlabeled primaries? Thanks, Kim Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA * This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ** ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: [IHCRG] Re: CMV antibody
Interestingly, This data sheet does not seem to list the clone of the CMV monoclonal antibody, or am I misreading it? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: ih...@googlegroups.com [mailto:ih...@googlegroups.com] On Behalf Of michael...@sickkids.ca Sent: Thursday, 21 May 2009 5:20 AM To: jtay...@meriter.com Cc: 'histonet@lists.utsouthwestern.edu'; 'ih...@googlegroups.com' Subject: [IHCRG] Re: CMV antibody See attached data sheet, we use pepsin enzyme digestion @ 37C for 30 minutes manually on FFPE For Ventana Users Protease I for 16 minutes Michael Ho, MLT Resource Technologist(See attached file: CMV-E.pdf) Immunopathology laboratory Division of Pathology DPLM The Hospital for Sick Children Toronto, Ontario Canada Tel#:416-813-5950 Fax: 416-813-5974 Taylor, Jean jtay...@meriter. com To Sent by: 'ih...@googlegroups.com' ih...@googlegroup ih...@googlegroups.com, s.com 'histonet@lists.utsouthwestern.edu ' histonet@lists.utsouthwestern.edu 2009-05-20 03:06 cc PM Subject [IHCRG] CMV antibody Please respond to jtay...@meriter.c om Hi Everyone, I'm wondering what antibody clone labs are using to detect Cytomegalovirus by IHC. Thank you! Jean Taylor, HT(ASCP) QIHC IHC Tech. Meriter Health Services Madison, WI 53715 * This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ** ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] cloudy formalin in tissue processor
Possible carry over of xylene from the flush program? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Christine I. Braaten Sent: Friday, 22 May 2009 2:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cloudy formalin in tissue processor Has anyone had a problem with their formalin getting cloudy in their tissue processor? We have a VIP tissue processor and just recently started having problems with cloudy formalin. Any suggestions would be appreciated. I checked the archives and couldn't find any articles on this particular subject. Thanks. CONFIDENTIALITY NOTICE: This electronic message, including any attachments, is for the sole use of the intended recipients and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by electronic mail and destroy all copies of the original message. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet * This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ** ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: [IHCRG] Re: CMV antibody
Michael, I have used both the Dako (Clones CCH2 + DDG9) and the Novocastra products (Clones 2 and 6) and they both stain the same. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: michael...@sickkids.ca [mailto:michael...@sickkids.ca] Sent: Friday, 22 May 2009 9:22 AM To: Tony Henwood Cc: histonet@lists.utsouthwestern.edu; ih...@googlegroups.com; jtay...@meriter.com Subject: Re: [IHCRG] Re: CMV antibody Hi Tony If you are interested I can email more information...touch base with me what are you using at your institute??? best wishes Michael Ho, MLT Resource Technologist Immunopathology laboratory Division of Pathology DPLM The Hospital for Sick Children Toronto, Ontario Canada Tel#:416-813-5950 Fax: 416-813-5974 Tony Henwood antho...@chw.edu .au To Sent by: michael...@sickkids.ca, ih...@googlegroup jtay...@meriter.com s.com cc histonet@lists.utsouthwestern.edu , ih...@googlegroups.com 05/21/2009 07:00 Subject PM[IHCRG] Re: CMV antibody Please respond to antho...@chw.edu. au Interestingly, This data sheet does not seem to list the clone of the CMV monoclonal antibody, or am I misreading it? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: ih...@googlegroups.com [mailto:ih...@googlegroups.com] On Behalf Of michael...@sickkids.ca Sent: Thursday, 21 May 2009 5:20 AM To: jtay...@meriter.com Cc: 'histonet@lists.utsouthwestern.edu'; 'ih...@googlegroups.com' Subject: [IHCRG] Re: CMV antibody See attached data sheet, we use pepsin enzyme digestion @ 37C for 30 minutes manually on FFPE For Ventana Users Protease I for 16 minutes Michael Ho, MLT Resource Technologist(See attached file: CMV-E.pdf) Immunopathology laboratory Division of Pathology DPLM The Hospital for Sick Children Toronto, Ontario Canada Tel#:416-813-5950 Fax: 416-813-5974 Taylor, Jean jtay...@meriter. com To Sent by: 'ih...@googlegroups.com' ih...@googlegroup ih...@googlegroups.com, s.com 'histonet@lists.utsouthwestern.edu ' histonet@lists.utsouthwestern.edu 2009-05-20 03:06 cc PM Subject [IHCRG] CMV antibody Please respond to jtay...@meriter.c om Hi Everyone, I'm wondering what antibody clone labs are using to detect Cytomegalovirus by IHC. Thank you! Jean Taylor, HT(ASCP) QIHC IHC Tech. Meriter Health Services Madison, WI 53715 * This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ** * This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ** ___ Histonet mailing list
[Histonet] re:cloudy formalin
Are you certain that it is formalin and not paraformaldehyde? The latter will get cloudy. Also, is there any way that your clearing solvent might be making its way in to the formalin solution? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Antibodies for Alport syndrome
Hi All, Has anyone had experience doing IHC on FFPE tissues with Collagen Type IV subunit antibodies? These are Collagen Type IV alpha 1, alpha 3 and alpha 5 chains. They are used for the diagnosis of Alport syndrome and all antibodies are from KAMIYA BIOMEDICAL COMPANY, Seatle. At present, it seems that only COL4A5 is working on FFPE tissues. I tried with (or without) various pre-treatment before the primary incubation with the dilution of 1:50 to 1:500. I am using BondMax autostainer and I also tried Decloaking chamber HIER with manual staining. I was wondering whether there would be other source for these antibodies. Thank you. Regards, Young Young Kwun Senior Hospital Scientist Dept. of Anatomical Pathology Concord Hospital Concord NSW 2139 Australia Tel)61-2-9767-6075 Fax)61-2-9767-8427 kw...@email.cs.nsw.gov.au ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Wholemount processing
Hello to all in histoland. My lab is looking to start processing whole prostate specimens and doing the wholemount process. Can anyone give me a starting point on how to proceed. I would need information on processing times, embedding and cutting equipment, staining information and the glass that you mount the specimens on. Any help in this matter would be greatly appreciated. Allison Scott HT(ASCP) Histology Supervisor LBJ Hospital Houston, Texas 77026 CONFIDENTIALITY NOTICE: If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail and any attachments from your computer system. To the extent the information in this e-mail and any attachments contain protected health information as defined by the Health Insurance Portability and Accountability Act of 1996 (HIPAA), PL 104-191; 45 CFR Parts 160 and 164; or Chapter 181, Texas Health and Safety Code, it is confidential and/or privileged. This e-mail may also be confidential and/or privileged under Texas law. The e-mail is for the use of only the individual or entity named above. If you are not the intended recipient, or any authorized representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail and its attachments is strictly prohibited. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
