RE: [Histonet] Path Course
Hi,
This looks really interesting. Do you know if there is the same type of
path courses/workshop for rodents pathology?
Regards
Jeanne
Jeanne Estabel, PhD
Scientific Manager
MGP Histology Operations Manager
Wellcome Trust Sanger Institute
Cambridge UK
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Patsy
Ruegg
Sent: 16 September 2010 17:55
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Path Course
Dear colleagues,
We just finalized the schedule for our fifth annual course. Many of you
have
already attended our previous courses. Feel free to forward this email
to
any interested colleagues.
Also, exhibitors are welcome for sponsorship opportunities. We have a
limited room this year, based on first come-first serve basis.
All information about schedule, location, time, faculty, exhibitors
prospectus, contact info, registration form, are available and
downloadable
online from the course's website at
www.pathlearning.com
Thanks,
Hadi
Hadi Yaziji, MD I Medical Director
Vitro Molecular Laboratories I www.vitromolecular.com
7480 SW 40th Street, Suite 700 Miami, FL 33155
Tel 305 267 7979 I Fax 786 513 0175
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 mailto:pru...@ihctech.net pru...@ihctech.net
web site http://www.ihctech.net 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
constitute a breach of civil or criminal law and is strictly prohibited.
If
you are NOT the intended recipient please contact the sender and dispose
of
this e-mail as soon as possible.
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
--
The Wellcome Trust Sanger Institute is operated by Genome Research
Limited, a charity registered in England with number 1021457 and a
company registered in England with number 2742969, whose registered
office is 215 Euston Road, London, NW1 2BE.
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] RE: Cutting, Processing, etc
i think that histonet is a primary educational group for all levels and any expert in our feild have asked these quistions one day befor and we should ask freely without any shame . i'm as begainner have learned alot from these little quistions. and i asked befor for name of antibodies and its use to be written to clear the information for all levels and no shame for that. --- On Thu, 9/16/10, Nails, Felton flna...@texaschildrens.org wrote: From: Nails, Felton flna...@texaschildrens.org Subject: [Histonet] RE: Cutting, Processing, etc To: Histonet@lists.utsouthwestern.edu Histonet@lists.utsouthwestern.edu Date: Thursday, September 16, 2010, 6:06 PM what is happening to our field?? -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Senn, Amy R Sent: Thursday, September 16, 2010 10:38 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Cutting, Processing, etc Hello Histoland! I have some questions about procedures in different histo labs and I'd like to have some 'backup' when people look at me like I'm crazy here... After embedding, you face (trim) your blocks, right? Do you take sections right from that same blade, or move/change your blade? How many of you 'soak' your blocks in water/softblock before cutting them? Do you put them on a cold plate/use ice and water, etc? Does this depend on the type of tissue, or do you treat them all the same? How often is freezy spray used in your lab, and where and when do you use it? How often do you rotate/change your reagents in your processors? Do you calculate this by how many blocks/days/weeks of use? Thank you so much for your input!! Amy Senn, HT Holy Spirit Hospital, Camp Hill PA Attention: This Message is intended only for the use of the individual or entity to which it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination or copying of this message or the taking of any action in reliance on the contents of this message is strictly prohibited. If you have received this message in error, please notify us immediately and destroy the original message. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- CONFIDENTIALITY NOTICE: The information in this e-mail may be confidential and/or privileged. If you are not the intended recipient or an authorized representative of the intended recipient, you are hereby notified that any review, dissemination, or copying of this e-mail and its attachments, if any, or the information contained herein is prohibited. If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail from your computer system. Thank you. == ___ 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] Rat heart valve histology trimming and sectioning reg
Dear Histonetters, Our pathologists are interested in rat heart valvular lesions. I am working on trimming and sectioning methods to get uniform and reproducible morphology of heart valves in paraffin sections. It seems that available literature dealing with this is limited. Requesting your valuable experience in this regard. Thanks a lot Abi jagannath ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Cassette Labeler
We have the Leica Cassette Labeler. It works just fine. I have looked at the Thermo Fisher Cassette Printer, which had a much smaller footprint than the Leica. I would be tempted to go with the Thermo if I have to replace my current labeler. These are expensive instruments but worth having. It would be appropriate for your workload. Plus, these cassette labelers can print a bar code, in addition to the pt. id and name. Laboratories can eliminate numbering errors by utilizing bar code scanners throughout the entire work flow; at grossing, processing and microtomy. Bar coding can also assist you with implementing LEAN processes. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of dianar...@aol.com Sent: Thursday, September 16, 2010 7:45 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cassette Labeler I work in a small lab and process approx 150 cassettes a day. We currently use a chemical resistant pen that works great. We write the number on top and the patient initials on the side. We are considering getting a cassette labeler. Is it really worth the expense for 150 blocks a day? Is it possible to enter information on the side of the cassette? Can you share some of your experience with different ones with me. ___ 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] Cassette Labeler
Thermo has a relatively low cost labeler for smaller quantities of cassette printing. The cost of the labeler versus the cost and potential for errors (hand writing) needs to be considered when making this kind of decision. You might want to do a study where you actually measure the time it takes to label one cassette. Make sure you start the clock from the time the tech actually prepares to label the cassette until they are finished and place the cassette down. Multiply that times the number of cassettes. Take that answer and multiply it times the tech salary plus benefits and you get the actual cost of hand labeling the cassettes. Steve -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of dianar...@aol.com Sent: Thursday, September 16, 2010 8:45 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cassette Labeler I work in a small lab and process approx 150 cassettes a day. We currently use a chemical resistant pen that works great. We write the number on top and the patient initials on the side. We are considering getting a cassette labeler. Is it really worth the expense for 150 blocks a day? Is it possible to enter information on the side of the cassette? Can you share some of your experience with different ones with me. ___ 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] Cassette Labeler
Good Morning, We process about that many blocks per day also, and I would say YES it is worth getting a cassette labeler. Leica/Surgipath has a small semi-automatic cassette labeler that we are in the process of buying and we LOVE it. It will print the accession# and the patient's name on the front of the cassette which complies with CAP's 2 pt identifiers. The automated part is you can set it to advance to the next acc # or advance the block or specimen numbers. Tonia Crook is our rep and her cell # is 803-237-9969 or call your Leica rep in your area. Happy cutting!!! Wanda WANDA G. SMITH, HTL(ASCP)HT Pathology Supervisor TRIDENT MEDICAL CENTER 9330 Medical Plaza Drive Charleston, SC 29406 843-847-4586 843-847-4296 fax This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of dianar...@aol.com Sent: Thursday, September 16, 2010 8:45 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cassette Labeler I work in a small lab and process approx 150 cassettes a day. We currently use a chemical resistant pen that works great. We write the number on top and the patient initials on the side. We are considering getting a cassette labeler. Is it really worth the expense for 150 blocks a day? Is it possible to enter information on the side of the cassette? Can you share some of your experience with different ones with me. ___ 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] caspase 3 ihc on rabbit tissue
Dear Friends, We are planning to perform active caspase 3 immunohistochemistry on FFPE rabbit cornea tissues. Most antibodies I have found are not working on rabbit. Any suggestion??? Thanks in advance... Dr. Necat Yilmaz Mersin University TURKEY __ ESET NOD32 Antivirus Akıllı Güvenlik tarafından sağlanan bilgiler, virüs imza veritabanı sürümü: 5458 (20100917) __ İleti ESET NOD32 Antivirus Akıllı Güvenlik tarafından denetlendi. http://www.nod32.com.tr ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Position in OH for 6-8 week coverage
Allied Search Partners has been retained to search for a Histotechnician or Histotechnologist who can remit their services to our client for 6-8 week coverage. The physician is looking for a qualified candidate to work ASAP. *Location of Laboratory:* Toledo, OH (local candidates please) *Environment:* Multi-specialty clinic serving the community with 108 physicians and 37 specialties. *Shift:* Full Time, Monday-Friday from 8am-5pm *Department:* Dermasurgery *Requirements: * HT ASCP Prior MOHS experienced preferred CLIA eligible to gross *Job Duties:* Grossing Assist physician Make permanent sections MOHS Histology Other related duties *Other:* * * This is not a temporary contract assignment. This is a 6-8 week coverage. You will be paid by the employer (our client), *so please submit your salary requirements with your resume* to be considered for this position. For more information and to schedule a phone interview with one of our recruiters please submit resume to aly...@alliedsearchpartners.com -- Alyssa Peterson, Director of Candidate Recruitment LinkedIN:http://www.linkedin.com/in/alyssapetersonasp Allied Search Partners T: 888.388.7571 F: 888.388.7572 www.alliedsearchpartners.com This email including its attachments is intended only for the confidential use of the individual to whom it is addressed. If you are not the intended recipient, any use, dissemination, distribution or copying of this message or its attachments is prohibited. If you have received this message in error, please notify us immediately, and delete this message and its attachments permanently from your system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Masson's trichrom - problem with nuclei staining
Hi Itai, I am interested to hear if you've resolved this problem. We use the same kit to stain mouse heart and embryo sections, PFA fixed. Our protocol is similar to the one you mentioned. I cannot get nuclei staining with this method either. The nuclei are well stained (ie black) up to the PMA/PTA step, but during that step the nuclear stain is completely removed. I cannot shorten the PMA/PTA step without negatively effecting the collagen stain. I have in our original protocol that the Weigert's working solution is good for one month, but I cannot recall if this is from Sigma's specification sheet or a personal observation from a lab member. However, from reading online some say it is good up to 4 months, others say it needs to be prepared fresh each time. I have tried fresh preparations with the same results. My instinct is that something is off - the staining is just not stable enough to withstand the subsequent acid steps in Masson's trichrome. Can an expert weigh in on this? Is there a way to strengthen nuclear staining from Weigert's? Sigma's formulas and usage recommendations are: Part A: 1% w/v certified Hematoxylin in ethanol Part B: 1.2% w/v Ferric chloride, 1% v/v Hydrochloric Acid Combine equal volumes Part A and Part B, stain sections for 5 minutes. Andrea Marion Graduate Student University of Illinois at Chicago amario3 /at/ uic /dot/ edu Itai Moshe itai.moshe @t mail.huji.ac.il Wed Sep 15 11:34:51 CDT 2010 Hi Histonet's I'm using Masson's trichrome to stain paraffin mice diaphragm fixed with PFA. I'm using this protocol: http://www.bcm.edu/mcb/rosenlab/index.cfm?pmid=12997 http://www.bcm.edu/mcb/rosenlab/index.cfm?pmid=12997With sigma masson's kit (#HT15) and Weigert's Iron Hematoxylin Set (#HT1079) The staining is beautiful, but i can't see the nuclei good enough. 1) Is there a way to enhance the nuclei staining ? (the nuclei is the only reason that im not using the simpler sirius red and fast green staining.) 2) What is the meaning for washing in running tap water washing, is it done by putting the slides in a jar with simple tap water for a few minutes ? Thank's Itai P.S By mistake I've post this before in another message with a wrong title. please respond to that message, so the title will be o.k for future archive searching. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Re: Masson's trichrom - problem with nuclei staining
I do Weigert's staining for 10 minutes and use it for a week at the most. I, too, have heard conflicting times about the stability of the Weigert's, but have the best results with using it no longer than a week. Deanna Rhoads HT (ASCP) From: Andrea Marion amar...@uic.edu To: histonet@lists.utsouthwestern.edu Cc: itai.mo...@mail.huji.ac.il Sent: Fri, September 17, 2010 10:31:16 AM Subject: [Histonet] Re: Masson's trichrom - problem with nuclei staining Hi Itai, I am interested to hear if you've resolved this problem. We use the same kit to stain mouse heart and embryo sections, PFA fixed. Our protocol is similar to the one you mentioned. I cannot get nuclei staining with this method either. The nuclei are well stained (ie black) up to the PMA/PTA step, but during that step the nuclear stain is completely removed. I cannot shorten the PMA/PTA step without negatively effecting the collagen stain. I have in our original protocol that the Weigert's working solution is good for one month, but I cannot recall if this is from Sigma's specification sheet or a personal observation from a lab member. However, from reading online some say it is good up to 4 months, others say it needs to be prepared fresh each time. I have tried fresh preparations with the same results. My instinct is that something is off - the staining is just not stable enough to withstand the subsequent acid steps in Masson's trichrome. Can an expert weigh in on this? Is there a way to strengthen nuclear staining from Weigert's? Sigma's formulas and usage recommendations are: Part A: 1% w/v certified Hematoxylin in ethanol Part B: 1.2% w/v Ferric chloride, 1% v/v Hydrochloric Acid Combine equal volumes Part A and Part B, stain sections for 5 minutes. Andrea Marion Graduate Student University of Illinois at Chicago amario3 /at/ uic /dot/ edu Itai Moshe itai.moshe @t mail.huji.ac.il Wed Sep 15 11:34:51 CDT 2010 Hi Histonet's I'm using Masson's trichrome to stain paraffin mice diaphragm fixed with PFA. I'm using this protocol: http://www.bcm.edu/mcb/rosenlab/index.cfm?pmid=12997 http://www.bcm.edu/mcb/rosenlab/index.cfm?pmid=12997With sigma masson's kit (#HT15) and Weigert's Iron Hematoxylin Set (#HT1079) The staining is beautiful, but i can't see the nuclei good enough. 1) Is there a way to enhance the nuclei staining ? (the nuclei is the only reason that im not using the simpler sirius red and fast green staining.) 2) What is the meaning for washing in running tap water washing, is it done by putting the slides in a jar with simple tap water for a few minutes ? Thank's Itai P.S By mistake I've post this before in another message with a wrong title. please respond to that message, so the title will be o.k for future archive searching. ___ 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] Re: Poor Weigerts Hematoxylin staining with Massons Trichrome
In general, we found the original/classic Weigerts iron hematoxylin stain to be weak and almost washed out of tissues after staining with Massons Trichrome. We no longer use the original formula, but a more concentrated modified formulation that is not differentiated out as much by the acidifed solutions found in Mass Tri. We also found this problem with Massons Trichrome kit components, where companies probably package the original method's solutions. We make up Weigerts fresh each time, and if it will last for a week for you, fine. but our work tends to be a one time staining with Mass Tri, then weeks before it was done again. The problem is: ferric chloride continues to oxidize the hematoxylin throughout over time, that week or longer, weakening the iron hematoxylin staining capabililty. This is discussed in Sheehan and Hrapchak Theory and Practice of Histotechnology. Acid decalcified bone presents even more of a challenge, since nuclei (DNA/RNA) in cells are hydrolyzed by acid decalcifiers, compromising staining of nuclei, a problem seen with routine HE staining. Deanna was correct on her assessment of this stain for best results. This modified Weigerts Hematoxylin was published in J of Histotechnology in a paper on Kreybergs stain on skin. The second Extra Strength Weigerts was found on Histonet years ago and I have not tried the latter. I suggest you see which one you prefer, as we use the first Modified Weigerts. Over the years, the modified gave us far superior nuclear staining with Massons Trichrome. Weigert's Iron Hematoxyin MODIFIED (found in J of Histotechnology in a method for Kreybergs stain on mouse skin). Solution A. 2% Hematoxylin in 95% ethanol Solution B. 62% Ferric Chloride 4 ml Distilled water 95 ml Hydrochloric acid 1 ml Mix equal amounts of Solution A and Solution B MIX FRESH JUST BEFORE USE AND DISCARD AFTER USE. Extra Strength Weigerts Iron Hematoxylin from Histonet (reference unknown, but supposedly from J of Histotechnology and method originated by Mabel Myli, Mayo Clinic) Solution A: Hematoxylin 10 g 95% ethanol 100 ml Solution B: 11.6 g Ferric Chloride 980 ml Distilled water 10 ml 25% hydrochloric acid Working Stain Solution Solution A 5 ml Solution B 25 ml Absolute Ethanol 20 ml Staining time for both of these formulations is 10 minutes, followed by rinsing for 10 min in running tap water (hematoxylin will be black) Gayle M. Callis HTL/HT/MT(ASCP) You Wrote: I do Weigert's staining for 10 minutes and use it for a week at the most. I, too, have heard conflicting times about the stability of the Weigert's, but have the best results with using it no longer than a week. Deanna Rhoads HT (ASCP) From: Andrea Marion amario3 http://lists.utsouthwestern.edu/mailman/listinfo/histonet @t uic.edu To: histonet http://lists.utsouthwestern.edu/mailman/listinfo/histonet @t lists.utsouthwestern.edu Cc: itai.moshe http://lists.utsouthwestern.edu/mailman/listinfo/histonet @t mail.huji.ac.il Sent: Fri, September 17, 2010 10:31:16 AM Subject: [Histonet] Re: Masson's trichrom - problem with nuclei staining Hi Itai, I am interested to hear if you've resolved this problem. We use the same kit to stain mouse heart and embryo sections, PFA fixed. Our protocol is similar to the one you mentioned. I cannot get nuclei staining with this method either. The nuclei are well stained (ie black) up to the PMA/PTA step, but during that step the nuclear stain is completely removed. I cannot shorten the PMA/PTA step without negatively effecting the collagen stain.In general, we found the original/ I have in our original protocol that the Weigert's working solution is good for one month, but I cannot recall if this is from Sigma's specification sheet or a personal observation from a lab member. However, from reading online some say it is good up to 4 months, others say it needs to be prepared fresh each time. I have tried fresh preparations with the same results. My instinct is that something is off - the staining is just not stable enough to withstand the subsequent acid steps in Masson's trichrome. Can an expert weigh in on this? Is there a way to strengthen nuclear staining from Weigert's? Sigma's formulas and usage recommendations are: Part A: 1% w/v certified Hematoxylin in ethanol Part B: 1.2% w/v Ferric chloride, 1% v/v Hydrochloric Acid Combine equal volumes Part A and Part B, stain sections for 5 minutes. Andrea Marion Graduate Student University of Illinois at Chicago amario3 /at/ uic /dot/ edu Itai Moshe itai.moshe @t mail.huji.ac.il Wed
RE: [Histonet] RE: Cutting, Processing, etc
My first reaction to the what is happening to our field, was WOW. It seemed unkind to me, as if they original poster should not have asked these questions. With further reading of the replies to this post, I am not so sure it was an unkind response, but one of potential shock and dismay to the idea that labs might not be producing the quality work that most of us employ on a daily basis. Amy, in answer to your questions, I will echo some of the sentiments that I have read here. 1. Facing of blocks. We use one blade to face blocks and another, new blade when we do our actual sectioning. In my case, I face as many as I can, knowing I am going to toss that knife when I am done facing. 2. Soaking of blocks. After facing my blocks, I will put them on a cold, damp, ice cube tray. This will achieve two purposes for me, a) to chill the block and b) to introduce moisture into the faced tissue. If I get a block that is particularly dry or hard (some calcified tissues for example), I will face them, put them face down on my waterbath and allow the hot water to penetrate into the tissue for 15-45 seconds, depending on the block. After cooling on the ice tray, they usually cut like butter for me. Typically, my blocks are not on the ice cubes for more than 15 minutes. As I cut some, I will rotate the blocks around the ice tray, adding more warm blocks for cooling. 3. Freeze spray. I hardly ever use the freeze spray. About the only time I find that I need it is if I have a particularly fatty tissue and it doesn't want to section. 4. Tissue processor changes. This is definitely something that is site specific. In our case, we do base it on volumes. If we have a small volume of our little biopsies, we might not change the machine weekly, but every two weeks. Generally our large specimen machine is changed weekly. Your mileage may vary! :o) Michelle -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of mohamed abd el razik Sent: Friday, September 17, 2010 5:38 AM To: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: Cutting, Processing, etc i think that histonet is a primary educational group for all levels and any expert in our feild have asked these quistions one day befor and we should ask freely without any shame . i'm as begainner have learned alot from these little quistions. and i asked befor for name of antibodies and its use to be written to clear the information for all levels and no shame for that. --- On Thu, 9/16/10, Nails, Felton flna...@texaschildrens.org wrote: From: Nails, Felton flna...@texaschildrens.org Subject: [Histonet] RE: Cutting, Processing, etc To: Histonet@lists.utsouthwestern.edu Histonet@lists.utsouthwestern.edu Date: Thursday, September 16, 2010, 6:06 PM what is happening to our field?? -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Senn, Amy R Sent: Thursday, September 16, 2010 10:38 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Cutting, Processing, etc Hello Histoland! I have some questions about procedures in different histo labs and I'd like to have some 'backup' when people look at me like I'm crazy here... After embedding, you face (trim) your blocks, right? Do you take sections right from that same blade, or move/change your blade? How many of you 'soak' your blocks in water/softblock before cutting them? Do you put them on a cold plate/use ice and water, etc? Does this depend on the type of tissue, or do you treat them all the same? How often is freezy spray used in your lab, and where and when do you use it? How often do you rotate/change your reagents in your processors? Do you calculate this by how many blocks/days/weeks of use? Thank you so much for your input!! Amy Senn, HT Holy Spirit Hospital, Camp Hill PA Attention: This Message is intended only for the use of the individual or entity to which it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination or copying of this message or the taking of any action in reliance on the contents of this message is strictly prohibited. If you have received this message in error, please notify us immediately and destroy the original message. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- CONFIDENTIALITY NOTICE: The information in this e-mail may be confidential and/or privileged. If you are not the intended recipient or an authorized representative of the intended
RE: [Histonet] RE: Cutting, Processing, etc
As I look through and monitor questions, it is apparent that our field is declining. These are very basic questions not about special stains or IHC stains but basic histology that should have been taught in histology 101. My fear is that as we get older and leave the field, who and what will be left to carry the torch. Those of you who ask, don't take offense to my thoughts but take action and pick up a book and read. You will improve yourself and the field. Just my thoughts, if I offended you it was not my intent. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of histot...@imagesbyhopper.com Sent: Friday, September 17, 2010 11:42 AM To: 'mohamed abd el razik'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Cutting, Processing, etc My first reaction to the what is happening to our field, was WOW. It seemed unkind to me, as if they original poster should not have asked these questions. With further reading of the replies to this post, I am not so sure it was an unkind response, but one of potential shock and dismay to the idea that labs might not be producing the quality work that most of us employ on a daily basis. Amy, in answer to your questions, I will echo some of the sentiments that I have read here. 1. Facing of blocks. We use one blade to face blocks and another, new blade when we do our actual sectioning. In my case, I face as many as I can, knowing I am going to toss that knife when I am done facing. 2. Soaking of blocks. After facing my blocks, I will put them on a cold, damp, ice cube tray. This will achieve two purposes for me, a) to chill the block and b) to introduce moisture into the faced tissue. If I get a block that is particularly dry or hard (some calcified tissues for example), I will face them, put them face down on my waterbath and allow the hot water to penetrate into the tissue for 15-45 seconds, depending on the block. After cooling on the ice tray, they usually cut like butter for me. Typically, my blocks are not on the ice cubes for more than 15 minutes. As I cut some, I will rotate the blocks around the ice tray, adding more warm blocks for cooling. 3. Freeze spray. I hardly ever use the freeze spray. About the only time I find that I need it is if I have a particularly fatty tissue and it doesn't want to section. 4. Tissue processor changes. This is definitely something that is site specific. In our case, we do base it on volumes. If we have a small volume of our little biopsies, we might not change the machine weekly, but every two weeks. Generally our large specimen machine is changed weekly. Your mileage may vary! :o) Michelle -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of mohamed abd el razik Sent: Friday, September 17, 2010 5:38 AM To: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: Cutting, Processing, etc i think that histonet is a primary educational group for all levels and any expert in our feild have asked these quistions one day befor and we should ask freely without any shame . i'm as begainner have learned alot from these little quistions. and i asked befor for name of antibodies and its use to be written to clear the information for all levels and no shame for that. --- On Thu, 9/16/10, Nails, Felton flna...@texaschildrens.org wrote: From: Nails, Felton flna...@texaschildrens.org Subject: [Histonet] RE: Cutting, Processing, etc To: Histonet@lists.utsouthwestern.edu Histonet@lists.utsouthwestern.edu Date: Thursday, September 16, 2010, 6:06 PM what is happening to our field?? -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Senn, Amy R Sent: Thursday, September 16, 2010 10:38 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Cutting, Processing, etc Hello Histoland! I have some questions about procedures in different histo labs and I'd like to have some 'backup' when people look at me like I'm crazy here... After embedding, you face (trim) your blocks, right? Do you take sections right from that same blade, or move/change your blade? How many of you 'soak' your blocks in water/softblock before cutting them? Do you put them on a cold plate/use ice and water, etc? Does this depend on the type of tissue, or do you treat them all the same? How often is freezy spray used in your lab, and where and when do you use it? How often do you rotate/change your reagents in your processors? Do you calculate this by how many blocks/days/weeks of use? Thank you so much for your input!! Amy Senn, HT Holy Spirit Hospital, Camp Hill PA Attention: This Message is intended only for the use of the individual or entity to which it is addressed, and may
RE: [Histonet] RE: Cutting, Processing, etc
My 2 cents is that she needed to convince someone this was how it is done! J Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 678-843-7376 - Phone 678-843-7831 - Fax -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Nails, Felton Sent: Friday, September 17, 2010 13:03 To: 'histot...@imagesbyhopper.com'; 'mohamed abd el razik'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Cutting, Processing, etc As I look through and monitor questions, it is apparent that our field is declining. These are very basic questions not about special stains or IHC stains but basic histology that should have been taught in histology 101. My fear is that as we get older and leave the field, who and what will be left to carry the torch. Those of you who ask, don't take offense to my thoughts but take action and pick up a book and read. You will improve yourself and the field. Just my thoughts, if I offended you it was not my intent. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of histot...@imagesbyhopper.com Sent: Friday, September 17, 2010 11:42 AM To: 'mohamed abd el razik'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Cutting, Processing, etc My first reaction to the what is happening to our field, was WOW. It seemed unkind to me, as if they original poster should not have asked these questions. With further reading of the replies to this post, I am not so sure it was an unkind response, but one of potential shock and dismay to the idea that labs might not be producing the quality work that most of us employ on a daily basis. Amy, in answer to your questions, I will echo some of the sentiments that I have read here. 1. Facing of blocks. We use one blade to face blocks and another, new blade when we do our actual sectioning. In my case, I face as many as I can, knowing I am going to toss that knife when I am done facing. 2. Soaking of blocks. After facing my blocks, I will put them on a cold, damp, ice cube tray. This will achieve two purposes for me, a) to chill the block and b) to introduce moisture into the faced tissue. If I get a block that is particularly dry or hard (some calcified tissues for example), I will face them, put them face down on my waterbath and allow the hot water to penetrate into the tissue for 15-45 seconds, depending on the block. After cooling on the ice tray, they usually cut like butter for me. Typically, my blocks are not on the ice cubes for more than 15 minutes. As I cut some, I will rotate the blocks around the ice tray, adding more warm blocks for cooling. 3. Freeze spray. I hardly ever use the freeze spray. About the only time I find that I need it is if I have a particularly fatty tissue and it doesn't want to section. 4. Tissue processor changes. This is definitely something that is site specific. In our case, we do base it on volumes. If we have a small volume of our little biopsies, we might not change the machine weekly, but every two weeks. Generally our large specimen machine is changed weekly. Your mileage may vary! :o) Michelle -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of mohamed abd el razik Sent: Friday, September 17, 2010 5:38 AM To: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: Cutting, Processing, etc i think that histonet is a primary educational group for all levels and any expert in our feild have asked these quistions one day befor and we should ask freely without any shame . i'm as begainner have learned alot from these little quistions. and i asked befor for name of antibodies and its use to be written to clear the information for all levels and no shame for that. --- On Thu, 9/16/10, Nails, Felton flna...@texaschildrens.org wrote: From: Nails, Felton flna...@texaschildrens.org Subject: [Histonet] RE: Cutting, Processing, etc To: Histonet@lists.utsouthwestern.edu Histonet@lists.utsouthwestern.edu Date: Thursday, September 16, 2010, 6:06 PM what is happening to our field?? -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Senn, Amy R Sent: Thursday, September 16, 2010 10:38 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Cutting, Processing, etc Hello Histoland! I have some questions about procedures in different histo labs and I'd like to have some 'backup' when people look at me like I'm crazy here... After embedding, you face (trim) your blocks, right? Do you take sections right from that same blade, or move/change your blade? How many of you 'soak' your blocks in water/softblock before cutting
RE: [Histonet] Rat heart valve histology trimming and sectioning reg
Assuming you are talking about the aortic valve Remove the heart and dissect away as much surrounding fat as you can. Leave some aorta attached. Make sure the heart is very well fixed in formalin. If the heart is soft and floppy rather than firm the next step wont go well. The next step is to remove most of the ventricles--you don't want to section through all that and it just gets in the way of proper orientation during embedding. There are two ways to trim off the ventricle. The critical thing is that your sectioning plane is perpendicular to the angle at which the aorta exits the heart. The classic way is to lay the heart in the anatomical position and use a straight blade, like a razor, to cut through it, guillotine-fashion along a line drawn between the lower margins of the atria and which is perpendicular to the angle at which the aorta exits the heart. I found that method to be not so reproducible so I came up with another way. Under a dissecting scope, I use my right hand and fine forceps to suspend the heart by the stub of the aorta. Gravity then automatically pulls the heart into almost the right position for the next step. Next I use my left hand and a straight back forceps to grasp the heart just below the atria. Then trim the aorta very close to the heart. Use your fine forceps to twiddle the angle that the heart sits in the straight forceps until you are looking directly down into the aortic sinus. You should be able to see the valves. Now take a number eleven scalpel and in one or two smooth strokes, run it along the edge of the straight forceps, separating the ventricles from the top of the heart. If you do it right you will have a sort of loaf shaped piece of tissue. If you lay it flat under the scope, the stub of the aorta will be pointing straight up and you will be able to see the valves. Embed the tissue so that the caudal side is at the bottom of the embedding mold. Throw away sections until you start to see the valves. Until you have cut a few and know what you are doing it is best to start saving sections early rather than later. I like to put three sections per slide and initially stain slides 1,3,4,7,9...etc, saving the rest for possible IHC. My prefered stain for lesions is the Modified Movat Pentachrome. It's kind of a pain to do but for lesion composition analysis it can't be beat. I start lesion analysis at the point where I can see all the leaflets. Stop when only the valve stems are visible. Anyway, it will take you a while before you get reproducible, nice cross sections. That first trimming away of the ventricles makes it or breaks it. Good luck and feel free to ask clarifying questions. Jerry Ricks Research Scientist University of Washington Department of Pathology Date: Fri, 17 Sep 2010 08:39:40 + To: histonet@lists.utsouthwestern.edu From: abija...@rediffmail.com Subject: [Histonet] Rat heart valve histology trimming and sectioning reg Dear Histonetters, Our pathologists are interested in rat heart valvular lesions. I am working on trimming and sectioning methods to get uniform and reproducible morphology of heart valves in paraffin sections. It seems that available literature dealing with this is limited. Requesting your valuable experience in this regard. Thanks a lot Abi jagannath ___ 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] RE: Cutting, Processing, etc
Felton, I would have to disagree with your assessment of the emails. Our field is very strong and is not in decline. Unfortunately, some supervisors around the country are relying on archaic methods and do not want to see or welcome change within the histo lab. We call ourselves professionals and yet not all of us are required to do continuing education? I read emails everyday and laugh at some of the bloviating that goes on inside this forum. I am glad the questions are asked, but I am also amazed at some of the responses that are shared with everyone. I choose to respond one on one with the person asking the question. Basic histology deals with didactics and this particular inquiry dealt more with OJT. There are many ways to get the same job done; are there more efficient ways? Probably, but this does not mean we all do our job the same way. I am not concerned about the future of Histotechnology. I embrace the opportunity to teach the young technicians about a field that sees a change almost daily. I am not here to offend either, but rather to defend an occupation that is as fascinating as it is frustrating. Respectfully, James Leroux, AAS, BA, HTL Histology Supervisor Petroglyph Pathology 640 Quantum Rd. Rio Rancho, NM 87124 (505) 924-0219 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Nails, Felton Sent: Friday, September 17, 2010 11:03 AM To: 'histot...@imagesbyhopper.com'; 'mohamed abd el razik'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Cutting, Processing, etc As I look through and monitor questions, it is apparent that our field is declining. These are very basic questions not about special stains or IHC stains but basic histology that should have been taught in histology 101. My fear is that as we get older and leave the field, who and what will be left to carry the torch. Those of you who ask, don't take offense to my thoughts but take action and pick up a book and read. You will improve yourself and the field. Just my thoughts, if I offended you it was not my intent. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of histot...@imagesbyhopper.com Sent: Friday, September 17, 2010 11:42 AM To: 'mohamed abd el razik'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Cutting, Processing, etc My first reaction to the what is happening to our field, was WOW. It seemed unkind to me, as if they original poster should not have asked these questions. With further reading of the replies to this post, I am not so sure it was an unkind response, but one of potential shock and dismay to the idea that labs might not be producing the quality work that most of us employ on a daily basis. Amy, in answer to your questions, I will echo some of the sentiments that I have read here. 1. Facing of blocks. We use one blade to face blocks and another, new blade when we do our actual sectioning. In my case, I face as many as I can, knowing I am going to toss that knife when I am done facing. 2. Soaking of blocks. After facing my blocks, I will put them on a cold, damp, ice cube tray. This will achieve two purposes for me, a) to chill the block and b) to introduce moisture into the faced tissue. If I get a block that is particularly dry or hard (some calcified tissues for example), I will face them, put them face down on my waterbath and allow the hot water to penetrate into the tissue for 15-45 seconds, depending on the block. After cooling on the ice tray, they usually cut like butter for me. Typically, my blocks are not on the ice cubes for more than 15 minutes. As I cut some, I will rotate the blocks around the ice tray, adding more warm blocks for cooling. 3. Freeze spray. I hardly ever use the freeze spray. About the only time I find that I need it is if I have a particularly fatty tissue and it doesn't want to section. 4. Tissue processor changes. This is definitely something that is site specific. In our case, we do base it on volumes. If we have a small volume of our little biopsies, we might not change the machine weekly, but every two weeks. Generally our large specimen machine is changed weekly. Your mileage may vary! :o) Michelle -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of mohamed abd el razik Sent: Friday, September 17, 2010 5:38 AM To: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: Cutting, Processing, etc i think that histonet is a primary educational group for all levels and any expert in our feild have asked these quistions one day befor and we should ask freely without any shame . i'm as begainner have learned alot from these little quistions. and i asked befor for name of antibodies and its use to be written to clear the
RE: [Histonet] RE: Cutting, Processing, etc
They do say that in the next 10 years 70% or so of us will be ret
ired. I am 30, and a rarity it seems. I think the main issue wi= th
people not picking up histology is because it is a fairly unknown
field.=I also think that with ASCP slowly moving towards having to
have a b= achelors degree the field is not updating itself. A college
graduate = starting in the world making the same pay they could
working at Walmart doe= sn't lend itself to that person learning or
staying in histology. I k= now the more experience you have the more
you make, but it still isn't enou= gh. I have been in the histo.
world since '98 and the pay scales real= ly haven't changed all that
much. I think the main solution would be = if you want college
educated, skilled HTs out there...pay them like a skill= ed college
graduate!! 35K a year for some places to start? How = do you pay
$400 a month in student loans and live with that salary? J= ust my 2
cents...
Oh and Amy...there are no stupid= questions!!! You go girl, learn
histology it's a great field. = Fair warning though...you will start
talking to yourself...it's a histo. th= ing =)
Sarah Goebel, B.A., HT (ASCP)
Histotechnician
XBiotech USA Inc.
8201 East Riverside Dr. Bldg= 4 Suite 100
Austin, Texas 78744
= em
(512)386-5= 107
Original Message
Subject: RE: [Histonet] RE: Cutting, Processing, etc
From: Nails, Felton [1]fln= a...@texaschildrens.org
Date: Fri, September 17, 2010 10:03 am
To: '[2]histot...@imagesbyhop= per.com' [3]histotech
@imagesbyhopper.com,
'mohamed abd el razik' [4]k8...@yahoo= .com,
[5]histo...@lists.utsout= hwestern.edu [6]
Histonet@lists.utsouthwestern.edu
As I look through and monitor questions, it is apparent that our field
is d= eclining. These are very basic questions not about special
stains or IHC st= ains but basic histology that should have been
taught in histology 101. My = fear is that as we get older and leave
the field, who and what will be left= to carry the torch. Those of you
who ask, don't take offense to my thought= s but take action and pick
up a book and read. You will improve yourself an= d the field.
Just my thoughts, if I offended you it was not my intent.
-Original Message-
From: [7]histonet= -boun...@lists.utsouthwestern.edu
[[8]mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
[9]histot...@= imagesbyhopper.com
Sent: Friday, September 17, 2010 11:42 AM
To: 'mohamed abd el razik'; [10]histo...@lists.utsouthwestern.edu
Subject: RE: [Histonet] RE: Cutting, Processing, etc
My first reaction to the what is happening to our field, was WOW. It
see= med unkind to me, as if they original poster should not have
asked these qu= estions. With further reading of the replies to this
post, I am not so sur= e it was an unkind response, but one of
potential shock and dismay to the i= dea that labs might not be
producing the quality work that most of us empl= oy on a daily basis.
Amy, in answer to your questions, I will echo some of the sentiments
that I= have read here.
1. Facing of blocks. We use one blade to face blocks and another, new
blad= e when we do our actual sectioning. In my case, I face as many
as I can, k= nowing I am going to toss that knife when I am done
facing.
2. Soaking of blocks. After facing my blocks, I will put them on a
cold, = damp, ice cube tray. This will achieve two purposes for me, a)
to chill th= e block and b) to introduce moisture into the faced
tissue. If I get a blo= ck that is particularly dry or hard (some
calcified tissues for example), I= will face them, put them face down
on my waterbath and allow the hot water= to penetrate into the tissue
for 15-45 seconds, depending on the block.
After cooling on the ice tray, they usually cut like butter for me.
Typically, my blocks are not on the ice cubes for more than 15
minutes. As= I cut some, I will rotate the blocks around the ice tray,
adding more war= m
blocks for cooling.
3. Freeze spray. I hardly ever use the freeze spray. About the only
time= I find that I need it is if I have a particularly fatty tissue
and it does= n't want to section.
4. Tissue processor changes. This is definitely something that is
site s= pecific. In our case, we do base it on volumes. If we have a
small volume= of our little biopsies, we might not change the
machine weekly, but ever= y two weeks. Generally our large specimen
machine is changed weekly.
Your mileage may vary! :o)
Michelle
-Original Message-
From: [11]histonet= -boun...@lists.utsouthwestern.edu
RE: [Histonet] RE: Cutting, Processing, etc
I echo Joyce's point. W/o all info, I hesitate to jump to conclusions. I was had an efficiency expert following me around for a week at the insistance of some administrators (it wasn't just me, but the entire lab)These are the types of silly questions these experts might ask, wondering if they can save 47 seconds out of the day. If anything is happening to our field, it might be the tampering by non-technicians in our technical duties in the name of stream-lining or keeping their tush's covered for their boss who is keeping her tush covered from ad nausium. It's a cynical Fridaybut the good news is that my Kindle is being delivered today! - Bill -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Weems, Joyce Sent: Friday, September 17, 2010 12:07 PM To: Nails, Felton; 'histot...@imagesbyhopper.com'; 'mohamed abd el razik'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Cutting, Processing, etc My 2 cents is that she needed to convince someone this was how it is done! J Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 678-843-7376 - Phone 678-843-7831 - Fax -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Nails, Felton Sent: Friday, September 17, 2010 13:03 To: 'histot...@imagesbyhopper.com'; 'mohamed abd el razik'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Cutting, Processing, etc As I look through and monitor questions, it is apparent that our field is declining. These are very basic questions not about special stains or IHC stains but basic histology that should have been taught in histology 101. My fear is that as we get older and leave the field, who and what will be left to carry the torch. Those of you who ask, don't take offense to my thoughts but take action and pick up a book and read. You will improve yourself and the field. Just my thoughts, if I offended you it was not my intent. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of histot...@imagesbyhopper.com Sent: Friday, September 17, 2010 11:42 AM To: 'mohamed abd el razik'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Cutting, Processing, etc My first reaction to the what is happening to our field, was WOW. It seemed unkind to me, as if they original poster should not have asked these questions. With further reading of the replies to this post, I am not so sure it was an unkind response, but one of potential shock and dismay to the idea that labs might not be producing the quality work that most of us employ on a daily basis. Amy, in answer to your questions, I will echo some of the sentiments that I have read here. 1. Facing of blocks. We use one blade to face blocks and another, new blade when we do our actual sectioning. In my case, I face as many as I can, knowing I am going to toss that knife when I am done facing. 2. Soaking of blocks. After facing my blocks, I will put them on a cold, damp, ice cube tray. This will achieve two purposes for me, a) to chill the block and b) to introduce moisture into the faced tissue. If I get a block that is particularly dry or hard (some calcified tissues for example), I will face them, put them face down on my waterbath and allow the hot water to penetrate into the tissue for 15-45 seconds, depending on the block. After cooling on the ice tray, they usually cut like butter for me. Typically, my blocks are not on the ice cubes for more than 15 minutes. As I cut some, I will rotate the blocks around the ice tray, adding more warm blocks for cooling. 3. Freeze spray. I hardly ever use the freeze spray. About the only time I find that I need it is if I have a particularly fatty tissue and it doesn't want to section. 4. Tissue processor changes. This is definitely something that is site specific. In our case, we do base it on volumes. If we have a small volume of our little biopsies, we might not change the machine weekly, but every two weeks. Generally our large specimen machine is changed weekly. Your mileage may vary! :o) Michelle -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of mohamed abd el razik Sent: Friday, September 17, 2010 5:38 AM To: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: Cutting, Processing, etc i think that histonet is a primary educational group for all levels and any expert in our feild have asked these quistions one day befor and we should ask freely without any shame . i'm as begainner have learned alot from these little quistions. and i asked befor for name of antibodies and its use to be written to clear the
[Histonet] unsubscribe
Jennifer M. Anderson, Scientist Halozyme Therapeutics, Inc. 11388 Sorrento Valley Road San Diego, CA 92121 858-704-8333 (office) jander...@halozyme.commailto:jander...@halozyme.com The information transmitted in this email is confidential and is intended only for the person(s) or entity to which it is addressed. Delivery of this message to any person other than the intended recipient(s) is not intended in any way to waive confidentiality or any applicable privilege. Any review, retransmission, dissemination or other use of, or taking of any action in reliance upon, this information by individuals or entities other than the intended recipient is prohibited by Halozyme and may be in violation of applicable laws. If you received this in error, please contact the sender and delete/destroy this email. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Rat heart valve histology trimming and sectioning reg
I just sent Abi a pdf of the publication he wanted. Joanna Barton and her colleagues developed a very nice way to use Histogel in a special way to optimize examination of rodent heart valves. After initial preparation/fixation they sliced the rat hearts using an acrylic rat heart matrix (slicing device) fronm Zivic Instruments. They ended up with three heart sections for further processing into paraffin. The photomicrographs showed excellent preservation of these very delicate but totally intact structures. I was very impressed at the quality of their work. Gayle Callis HTL/HT/MT(ASCP) -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of JR R Sent: Friday, September 17, 2010 11:16 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Rat heart valve histology trimming and sectioning reg Assuming you are talking about the aortic valve Remove the heart and dissect away as much surrounding fat as you can. Leave some aorta attached. Make sure the heart is very well fixed in formalin. If the heart is soft and floppy rather than firm the next step wont go well. The next step is to remove most of the ventricles--you don't want to section through all that and it just gets in the way of proper orientation during embedding. There are two ways to trim off the ventricle. The critical thing is that your sectioning plane is perpendicular to the angle at which the aorta exits the heart. The classic way is to lay the heart in the anatomical position and use a straight blade, like a razor, to cut through it, guillotine-fashion along a line drawn between the lower margins of the atria and which is perpendicular to the angle at which the aorta exits the heart. I found that method to be not so reproducible so I came up with another way. Under a dissecting scope, I use my right hand and fine forceps to suspend the heart by the stub of the aorta. Gravity then automatically pulls the heart into almost the right position for the next step. Next I use my left hand and a straight back forceps to grasp the heart just below the atria. Then trim the aorta very close to the heart. Use your fine forceps to twiddle the angle that the heart sits in the straight forceps until you are looking directly down into the aortic sinus. You should be able to see the valves. Now take a number eleven scalpel and in one or two smooth strokes, run it along the edge of the straight forceps, separating the ventricles from the top of the heart. If you do it right you will have a sort of loaf shaped piece of tissue. If you lay it flat under the scope, the stub of the aorta will be pointing straight up and you will be able to see the valves. Embed the tissue so that the caudal side is at the bottom of the embedding mold. Throw away sections until you start to see the valves. Until you have cut a few and know what you are doing it is best to start saving sections early rather than later. I like to put three sections per slide and initially stain slides 1,3,4,7,9...etc, saving the rest for possible IHC. My prefered stain for lesions is the Modified Movat Pentachrome. It's kind of a pain to do but for lesion composition analysis it can't be beat. I start lesion analysis at the point where I can see all the leaflets. Stop when only the valve stems are visible. Anyway, it will take you a while before you get reproducible, nice cross sections. That first trimming away of the ventricles makes it or breaks it. Good luck and feel free to ask clarifying questions. Jerry Ricks Research Scientist University of Washington Department of Pathology Date: Fri, 17 Sep 2010 08:39:40 + To: histonet@lists.utsouthwestern.edu From: abija...@rediffmail.com Subject: [Histonet] Rat heart valve histology trimming and sectioning reg Dear Histonetters, Our pathologists are interested in rat heart valvular lesions. I am working on trimming and sectioning methods to get uniform and reproducible morphology of heart valves in paraffin sections. It seems that available literature dealing with this is limited. Requesting your valuable experience in this regard. Thanks a lot Abi jagannath ___ 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 __ Information from ESET Smart Security, version of virus signature database 5458 (20100917) __ The message was checked by ESET Smart Security. http://www.eset.com __ Information from ESET Smart Security, version of virus signature database 5458 (20100917) __ The message was checked by ESET Smart Security. http://www.eset.com
[Histonet] Autofluorescence problem on atheroma - TUNEL staining
Hi, Autofluorescence in atheroma is a well known problem for immunofluorescence staining. I would love to hear input about how to reduce it to a minimum (if possible!). So far, we are trying to detect apoptotic cells in atheroma by TUNEL staining (Roche, TMRred) and we found the interpretation very difficult as some area of the tissue show red dots, tat could be considered as apoptotic bodies. We are working with frozen sections, fixing with Formalin, quenching with Glycine etc... -I was wondering if acetone fixation (or other fixation methods) could decrease atheroma autofluorescence by extracting lipids and others materials? -Also, I am not sure that acetone fixation is recommended for TUNEL staining? Did anybody try? -Are there around any approved ways to decrease autofluorescence in atheroma? (I can not find anything!) Thanks! Delphine Delphine Eberlé PhD UCSF Department of Vascular Surgery VA Medical Center - NCIRE Building 2 - room 410 4150 Clement Street San Francisco, CA 94121, USA Tel: 415 221 4810 ext.2984 Cell: 857 453 0821 delphine.ebe...@ucsfmedctr.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] UKAIH- Temp or Traveling Histotech needed asap!!!
Calling all histotechs who need a few extra hours!!! A very good friend of mine is in need of a temp or traveling histotech to cover the histology duties for 2 weeks... potentially one month. STARTING NOW! Low volume (40-60 blocks) For more details call Shawn 707-463-7301 Clinical Lab Supervisor or the histology lab asst Pam at 707-462-3111 x1480 The pathologist is very sweet, I love him! I have done work for him before. I live 2 hours away, so there's no way I can commute and manage my lab as well. Hours are 8am to 430pm... (or less if you can get it done faster) Maria Katleba MS HT(ASCP) Pathology Dept. Mgr Queen of the Valley Medical Center 1000 Trancas Street Napa CA 94558 (707) 252-4411 x3689 direct (707) 226-4385 pager (707) 294-9229 cell- anytime Notice from St. Joseph Health System: Please note that the information contained in this message may be privileged and confidential and protected from disclosure. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Thomas Crowell is out of the office.
I will be out of the office starting 09/17/2010 and will not return until 09/28/2010. Please contact Kelly Miner at 617-871-5122 if you have any questions regarding clinical trial samples. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Ventana Ultraview Detection
Hello Histonetters, We are in the process of selling some of our Ventana Ultraview Detection kits. The kits still have a long shelf life and have not been registered. If you are interested in purchasing one of more of these kits please email me with questions and or contact info. Thanks, Timothy Higgins, HT (ASCP) QIHC Histology Manager APA Amarillo, TX ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] RE: Cutting, Processing, etc
Hi James, I would take it a step farther with the continuing ed. I think it's beyond the supervisors it gets up into lab administration (clinical lab world). I personally know of a group of great folks that work hard and run a quality service. In the last 3 years they've had a major drop off in their continuing ed. And it, of course, is tied to the budget. Unfortunately, in this case (my view) those making the money decisions are missing the value. It seems they're unwilling to make the investment. I fear that in 5 years or less (if it continues) this service will suffer. I suspect there are other folks out there in the same boat. My hat is off to everyone out there working hard in our field and to the enlightened administrators and physicians that advocate continuing ed. Have a great weekend. Tom Jasper Thomas Jasper HT (ASCP) BAS Histology Supervisor Central Oregon Regional Pathology Services Bend, Oregon 97701 541/693-2677 tjas...@copc.net -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of james leroux Sent: Friday, September 17, 2010 10:55 AM To: 'Nails, Felton'; histot...@imagesbyhopper.com; 'mohamed abd el razik'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Cutting, Processing, etc Felton, I would have to disagree with your assessment of the emails. Our field is very strong and is not in decline. Unfortunately, some supervisors around the country are relying on archaic methods and do not want to see or welcome change within the histo lab. We call ourselves professionals and yet not all of us are required to do continuing education? I read emails everyday and laugh at some of the bloviating that goes on inside this forum. I am glad the questions are asked, but I am also amazed at some of the responses that are shared with everyone. I choose to respond one on one with the person asking the question. Basic histology deals with didactics and this particular inquiry dealt more with OJT. There are many ways to get the same job done; are there more efficient ways? Probably, but this does not mean we all do our job the same way. I am not concerned about the future of Histotechnology. I embrace the opportunity to teach the young technicians about a field that sees a change almost daily. I am not here to offend either, but rather to defend an occupation that is as fascinating as it is frustrating. Respectfully, James Leroux, AAS, BA, HTL Histology Supervisor Petroglyph Pathology 640 Quantum Rd. Rio Rancho, NM 87124 (505) 924-0219 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Nails, Felton Sent: Friday, September 17, 2010 11:03 AM To: 'histot...@imagesbyhopper.com'; 'mohamed abd el razik'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Cutting, Processing, etc As I look through and monitor questions, it is apparent that our field is declining. These are very basic questions not about special stains or IHC stains but basic histology that should have been taught in histology 101. My fear is that as we get older and leave the field, who and what will be left to carry the torch. Those of you who ask, don't take offense to my thoughts but take action and pick up a book and read. You will improve yourself and the field. Just my thoughts, if I offended you it was not my intent. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of histot...@imagesbyhopper.com Sent: Friday, September 17, 2010 11:42 AM To: 'mohamed abd el razik'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Cutting, Processing, etc My first reaction to the what is happening to our field, was WOW. It seemed unkind to me, as if they original poster should not have asked these questions. With further reading of the replies to this post, I am not so sure it was an unkind response, but one of potential shock and dismay to the idea that labs might not be producing the quality work that most of us employ on a daily basis. Amy, in answer to your questions, I will echo some of the sentiments that I have read here. 1. Facing of blocks. We use one blade to face blocks and another, new blade when we do our actual sectioning. In my case, I face as many as I can, knowing I am going to toss that knife when I am done facing. 2. Soaking of blocks. After facing my blocks, I will put them on a cold, damp, ice cube tray. This will achieve two purposes for me, a) to chill the block and b) to introduce moisture into the faced tissue. If I get a block that is particularly dry or hard (some calcified tissues for example), I will face them, put them face down on my waterbath and allow the hot water to penetrate into the tissue for 15-45 seconds, depending on the block. After
[Histonet] Out of Office Reply
I will be out the office the week of September 20-24. I will return on Monday, September 27th. If you need laboratory assistance please call 785-273-2788 ext. 322. Thanks. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Is anybody using TBS ATP1 - 120 tissue processor?
Hi all, Would you please let me know off line what do you think about this unit? macve...@usc.edu Thank you very much in advance Michelle Aloni MS HTL ASCP ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Histology Training Specialist in the Chicago Area!!
Good Afternoon! Please contact me directly for more information on this position or to learn more about our other opportunities. Have a great weekend! Histology Training Specialist The Company: Our client is a leading developer and producer of innovative high-tech precision optics systems for the analysis of microstructures. As one of the market leaders in each of the fields of Microscopy, Confocal Laser Scanning Microscopy, Imaging Systems, Specimen Preparation and Medical Equipment. Comprising nine manufacturing facilities in seven countries, sales and service companies in 20 countries and an international network of dealers, the company is represented in over 100 countries. The Opportunity: The company currently has an opening for a Histology Training Specialist. All applicants must not be adverse to travel, as this is a position that may require you to travel when necessary. Base: $55-60k Other: Full benefits - 401k program/matching Primary Responsibilities: The primary responsibility of this role will be to provide technical phone support by answering questions, troubleshooting problems, logging and closing complaint files and escalating major issues to appropriate company personnel. This role will also provide technical training on specified products in the company's newly constructed state-of-the-art Customer Support Laboratory. Training programs are designed for small groups to ensure maximum customer learning and satisfaction. Additional Responsibilities: - Provide product and applications phone support to end-users, field personnel and dealers for all product lines - Log all calls into Customer Support Database - Participate in development of training materials and conduct classes and labs for customers, employees and others as needed - Serve as technical liaison to Customer Service/Field Service/Product Management departments Education and Experience Required: Ability to interact with various people in a calm and positive fashion and the ability to effectively communicate information to groups of participants is required. Experience with data entry, MS Office programs (PowerPoint, Lotus Notes, Word) is also required. HT/HTL/QIHC (ASCP) is helpful but not required. Matt Ward Account Executive Personify 201 Shannon Oaks Circle, Suite 101 Cary, North Carolina 27511 (Tel) 800.875.6188 direct ext 103 (Fax) 919.460.0642 http://www.personifysearch.com/ www.personifysearch.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] In search of CRO specializing in volumetric determination via image analysis on serial sections.
Looking for a CRO experienced in determining the volume of an irregularly shaped object from serial sections. Automation experience a plus. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
