[Histonet] Eosinophilic new bone formation
We have always noticed, at least in the middle/inner ear, that newly deposited bone stains darker than mature bone, both with HE and toluidine blue. Is this increased eosinophilic quality due to a lack of mineralization and therefore higher density of osteoid components in the new bone or some other difference in composition of the osteoid? The contrast is quite striking when we observe bone remodeling due to middle ear infections, so I wanted to be able to offer an accurate explanation of why that is... Patricia Keller Sr. Research Tech/Core Histologist Washington University School of Medicine Department of Otolaryngology 4566 Scott Ave Campus Box 8115 St. Louis, Mo 63110 314-747-7166 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 have received this email in error please notify the system manager. This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this e-mail. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Tissue Processors
Sahura is still the best buy (for me). René J. --- On Thu, 2/17/11, Joe Nocito jnoc...@satx.rr.com wrote: From: Joe Nocito jnoc...@satx.rr.com Subject: [Histonet] Tissue Processors To: Histonet histonet@lists.utsouthwestern.edu Date: Thursday, February 17, 2011, 8:55 PM Greetings all, if you had to purchase new tissue processors, which one would you choose? Microwave technology is out of the question. Are Sakura's still a good buy? We've tried the Leica Peloris and the Shandon Pathcenters. Thanks for your help Joe ___ 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] Tissue Processors
I would probably have gone along with you on this Rene but since we demo'ed, and subsequently purchased, the Peloris my opinion has changed Ronnie Houston Anatomic Pathology Manager Nationwide Children's Hospital Columbus OH 43205 (614) 722 5450 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Friday, February 18, 2011 8:33 AM To: Histonet; Joe Nocito Subject: Re: [Histonet] Tissue Processors Sahura is still the best buy (for me). René J. --- On Thu, 2/17/11, Joe Nocito jnoc...@satx.rr.com wrote: From: Joe Nocito jnoc...@satx.rr.com Subject: [Histonet] Tissue Processors To: Histonet histonet@lists.utsouthwestern.edu Date: Thursday, February 17, 2011, 8:55 PM Greetings all, if you had to purchase new tissue processors, which one would you choose? Microwave technology is out of the question. Are Sakura's still a good buy? We've tried the Leica Peloris and the Shandon Pathcenters. Thanks for your help Joe ___ 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 - Confidentiality Notice: The following mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. The recipient is responsible to maintain the confidentiality of this information and to use the information only for authorized purposes. If you are not the intended recipient (or authorized to receive information for the intended recipient), you are hereby notified that any review, use, disclosure, distribution, copying, printing, or action taken in reliance on the contents of this e-mail is strictly prohibited. If you have received this communication in error, please notify us immediately by reply e-mail and destroy all copies of the original message. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Eosinophilic new bone formation
In decalcified sections of bone, yes the osteoid or dense unmineralized collagen matrix (mostly type I) will stain darker than mature native demineralized bone. Even though the mature bone has been demineralized, it is still more densely compact as compared to the newly formed bone that has not begun mineralization and . In resin embedded undemineralized sections of bone, the contrast is exactly opposite when staining with Von Kossa and counterstaining with MacNeal's tetrachrome and similar to decalcified sections in a Goldner's trichrome where the acid fuchsin stains osteoid darker than the light green does the mineralized bone. So the answer is yes, tissue density is what plays the major role in contrast staining and stain intensity. Jack On Feb 18, 2011, at 6:45 AM, Keller, Pat kell...@ent.wustl.edu wrote: We have always noticed, at least in the middle/inner ear, that newly deposited bone stains darker than mature bone, both with HE and toluidine blue. Is this increased eosinophilic quality due to a lack of mineralization and therefore higher density of osteoid components in the new bone or some other difference in composition of the osteoid? The contrast is quite striking when we observe bone remodeling due to middle ear infections, so I wanted to be able to offer an accurate explanation of why that is... Patricia Keller Sr. Research Tech/Core Histologist Washington University School of Medicine Department of Otolaryngology 4566 Scott Ave Campus Box 8115 St. Louis, Mo 63110 314-747-7166 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 have received this email in error please notify the system manager. This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this 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
RE: [Histonet] Tissue Processors
Pathcenters in my opinion don't last as long? Sarah Goebel, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Thursday, February 17, 2011 7:56 PM To: Histonet Subject: [Histonet] Tissue Processors Greetings all, if you had to purchase new tissue processors, which one would you choose? Microwave technology is out of the question. Are Sakura's still a good buy? We've tried the Leica Peloris and the Shandon Pathcenters. Thanks for your help Joe ___ 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] Microwave Usage, Monitoring procedure
Hello to all in histoland. I am in need of a procedure for the usgae, monitoring and container venting of the mcrowave. I have a procedure that I wrote several years ago, but my boss wants it updated. Any help will be appreciated. Allison Scott HT(ASCP) Histology Supervisor LBJ Hospital Houston, Texas 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
[Histonet] RE: Histonet Digest, Vol 87, Issue 33
We are also fans of the Leica ASP 300 processor. Diana G. Goodwin, BS, HT(ASCP)QIHC Department of Pathology Robert Wood Johnson University Hospital at Hamilton Hamilton, NJ 08610 609-631-6996 dgood...@rwjuhh.edu -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of histonet-requ...@lists.utsouthwestern.edu Sent: Friday, February 18, 2011 1:05 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 87, Issue 33 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. Tissue Processors (Joe Nocito) 2. RE: Tissue Processors (Langenberg, Stacey) 3. IHC pretreatment with NaOH/H2O2 (Neil M. Fournier) 4. Eosinophilic new bone formation (Keller, Pat) 5. Re: Tissue Processors (Rene J Buesa) 6. RE: Tissue Processors (Nails, Felton) 7. RE: Tissue Processors (Houston, Ronald) 8. Re: Eosinophilic new bone formation (Jack Ratliff) 9. Histotechnologist with a FL HTL license for Fort Myers, FL (Brian- Prometheus) 10. RE: Tissue Processors (sgoe...@mirnarx.com) 11. Microwave Usage, Monitoring procedure (Scott, Allison D) -- Message: 1 Date: Thu, 17 Feb 2011 19:55:53 -0600 From: Joe Nocito jnoc...@satx.rr.com Subject: [Histonet] Tissue Processors To: Histonet histonet@lists.utsouthwestern.edu Message-ID: 48ABF837D7084DCCB45CADF5F2B3F20A@JoePC Content-Type: text/plain; charset=iso-8859-1 Greetings all, if you had to purchase new tissue processors, which one would you choose? Microwave technology is out of the question. Are Sakura's still a good buy? We've tried the Leica Peloris and the Shandon Pathcenters. Thanks for your help Joe -- Message: 2 Date: Thu, 17 Feb 2011 19:33:13 -0700 From: Langenberg, Stacey stacey.langenb...@ucdenver.edu Subject: RE: [Histonet] Tissue Processors To: Joe Nocito jnoc...@satx.rr.com, Histonet histonet@lists.utsouthwestern.edu Message-ID: 1f70fcbb6d4ec549b2adf69b9f9eac034e6b03f...@steamboat.ucdenver.pvt Content-Type: text/plain; charset=us-ascii Joe, We are really impressed with the Leica ASP 300. We have a second one on the way. Stacey People are not an interruption of our business. People are our business. Stacey Langenberg HT (ASCP) QIHC Laboratory Manager Histology/IF CU Dermatopathology Consultants 1999 N. Fitzsimons Pkwy Suite 120 Aurora, CO 80045 Lab-720-859-3559 Fax- 303-344-0789 Office- 303-577-2303 Cell-970-405-7742 From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Joe Nocito [jnoc...@satx.rr.com] Sent: Thursday, February 17, 2011 6:55 PM To: Histonet Subject: [Histonet] Tissue Processors Greetings all, if you had to purchase new tissue processors, which one would you choose? Microwave technology is out of the question. Are Sakura's still a good buy? We've tried the Leica Peloris and the Shandon Pathcenters. Thanks for your help Joe ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Message: 3 Date: Fri, 18 Feb 2011 00:53:30 -0500 From: Neil M. Fournier neil.fourn...@yale.edu Subject: [Histonet] IHC pretreatment with NaOH/H2O2 To: histonet@lists.utsouthwestern.edu Message-ID: 20110218005330.8h3p3wjcu84gc...@www.mail.yale.edu Content-Type: text/plain; charset=ISO-8859-1 I recently came across a protocol that was conducting IHC staining for pSTAT3 on free-floating fixed rat brain sections. The protocol stated that the tissue was placed in 1% NaOH and 1% H2O2 in H2O for 20 min, 0.3% glycine for 10 min, and 0.03% sodium dodecyl sulfate for 10 min. Could someone please explain to me the rationale for using NaOH and H2O2 pretreatment, as well as the additional steps in this procedure? My rationale may be incorrect but I would be concerned that the combination of NaOH and H2O2 might produce a too much O2 bubbling that could harm the tissue. I believe glycine is typically used for reduce autofluoresence since it binds to free aldehydes but this protocol was utilizing a cobolt/nickel enhanced-DAB reaction and I am unaware of any empirical study that has shown that glycine pretreatment improves peroxidase immunostaining. Finally, SDS seems like a fairly harsh detergent (although the
[Histonet] Re: IHC pretreatment with NaOH/H2O2
Dear Neil, I was able to find a protocol like you described published where they had done a regular IHC protocol on free-floating tissue sections for a-MSH antibody using 0.3% H2O2, normal serum block, etc. They go on to describe that for P-STAT3, the tissue needed to be pretreated with 1% NaOH and 1%H2O2 in water for 20 minutes, 0.3% glycine for 10 minutes, and 0.03% SDS for 10 minutes. After that it looks like a standard protocol. Here's the URL for that paper: http://endo.endojournals.org/cgi/reprint/144/5/2121?ijkey=23f435f1cf61629953465217ca752dd73df366e9 They don't say why this was necessary, but it might be worth trying to track down one of the authors and ask them. It seems like the sodium hydroxide and hydrogen peroxide serves as an oxidizer somehow. Since they are not doing IF, maybe the glycine is needed to bind with glycine receptors in the brain prior to staining? And I suspect the SDS is needed for permeabilization, even though they are using a sectioned sample. The right detergent can make all the difference. I would really be interested in knowing what the rationale is, but it seems like they figured out what physiologic changes needed to happen for this antibody to work in brain. If you do have a conversation with the authors, please report back here because my curiosity is now at high roar. Best wishes, Teri Johnson, HT(ASCP)QIHC Head, Histology and Electron Microscopy Stowers Institute for Medical Research Kansas City, MO ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Re: IHC pretreatment with NaOH/H2O2
Neil, When I read the paper in question, it reminded me of the pretreatments one would use in an In Situ Hybridization protocol. I have used this protocol in the past and it doesn't seem to harm the tissue as long as the tissue has been adequately perfused. We use the pSTAT3 #9131 (Cell Signaling) with 40um free-floating rat brainstem sections that have been perfused with 4% paraformaldehyde . After successfully reproducing their protocol, I decided to try and incorporate my own protocol for this particular antibody. The only pretreatment that I use is a cocktail of Methanol (cold) + 1% H2O2 for 20 minutes., TBST Rinses, Blocking step - Rodent Block R, Primary - pSTAT3 (1:50 - 1:100) - overnight, TBST Rinses, Alexa Fluor secondary - 2hrs., TBST Rinses, mount coverslip with ProLong Gold (Invitrogen). **The #9131 spec sheet notes that you must use Methanol as a pretreatment step for this anitbody. *** I am often required to use this antibody in double labeling experiments with tract tracing beads that are sensitive to heat. HIER is out of the question, but the methanol/H2O2 pretreatment appears to be adequate for this antibody and doesn't affect the beads. Regards, Tina Montina J. Van Meter Lab Manager Pennington Biomedical Research Center Autonomic Neuroscience -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Johnson, Teri Sent: Friday, February 18, 2011 1:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: IHC pretreatment with NaOH/H2O2 Dear Neil, I was able to find a protocol like you described published where they had done a regular IHC protocol on free-floating tissue sections for a-MSH antibody using 0.3% H2O2, normal serum block, etc. They go on to describe that for P-STAT3, the tissue needed to be pretreated with 1% NaOH and 1%H2O2 in water for 20 minutes, 0.3% glycine for 10 minutes, and 0.03% SDS for 10 minutes. After that it looks like a standard protocol. Here's the URL for that paper: http://endo.endojournals.org/cgi/reprint/144/5/2121?ijkey=23f435f1cf6162 9953465217ca752dd73df366e9 They don't say why this was necessary, but it might be worth trying to track down one of the authors and ask them. It seems like the sodium hydroxide and hydrogen peroxide serves as an oxidizer somehow. Since they are not doing IF, maybe the glycine is needed to bind with glycine receptors in the brain prior to staining? And I suspect the SDS is needed for permeabilization, even though they are using a sectioned sample. The right detergent can make all the difference. I would really be interested in knowing what the rationale is, but it seems like they figured out what physiologic changes needed to happen for this antibody to work in brain. If you do have a conversation with the authors, please report back here because my curiosity is now at high roar. Best wishes, Teri Johnson, HT(ASCP)QIHC Head, Histology and Electron Microscopy Stowers Institute for Medical Research Kansas City, MO ___ 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] Information Needed
Hi, I need information for purchasing a good vacuum with proper biological filtration for cleaning a cryostat. We are not buying a new cryostat and I do know of several with a decontamination feature, it is not in the budget for the next two years. Thanks, Pamela A Marcum Anatomic Pathology Manager University of Arkansas for Medical Sciences 4301 W Markham St Slot 502 Little Rock AR 72205 Office: 501-686-5941 Pager: 501-395-1943 Fax: 501-686-7151 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] Job Descriptions
Hello Histoland, I was wondering if anyone out there may have their job decriptions for histologists, lab assistants, supervisor and/or medical director available and would be willing to share. I am currently revamping mine and am struggling with some of the verbiage. Thank you all and have a fantastic weekend! Cristi ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] LKB Bromma 7800 knifemaker
Yes, I know this is ancient. But does anyone know of a service company that can repair this machine? Ours works, sort of, but it's very frustrating. Thanks and have a good weekend, Kathleen Roberts Principal Lab Technician Neurotoxicology Labs Molecular Pathology Facility Core Dept of Pharmacology Toxicology Rutgers, the State University of NJ 41 B Gordon Road Piscataway, NJ 08854 (732) 445-6914 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Job Opening
We are looking for a certified HT or HTL to join our pathology lab in Manitowoc, WI. We have a currently 2nd shift position open. We have excellent benefits, state-of-the-are lab (less than 3 months old) and a great team to work with. We will assist with relocation. Please contact me if you have any questions. Michael Hillmer PHR HR Coordinator Dermatology Associates of Wisconsin Phone: (920)683-5278 Fax: (920)686-9674 Cell: (920)860-6360 The materials and information in this e-mail are confidential and may contain Protected Health Information covered under the HIPAA Privacy Rule. If you are not the intended recipient, be advised that any unauthorized use, disclosure, copying, distribution, or action taken in reliance on the contents of this information is strictly forbidden by law. If you have received this e-mail in error, please notify me by reply e-mail and then delete this message. Do not pass any of this information to anyone else. Thank you for your cooperation. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Information Needed
Pamela We use a hepa filtered vacuum from Marmed, Inc. (440) 572-5175 LiZ Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Marcum, Pamela A Sent: Friday, February 18, 2011 1:08 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Information Needed Hi, I need information for purchasing a good vacuum with proper biological filtration for cleaning a cryostat. We are not buying a new cryostat and I do know of several with a decontamination feature, it is not in the budget for the next two years. Thanks, Pamela A Marcum Anatomic Pathology Manager University of Arkansas for Medical Sciences 4301 W Markham St Slot 502 Little Rock AR 72205 Office: 501-686-5941 Pager: 501-395-1943 Fax: 501-686-7151 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
