[Histonet] Protocol for fluorescence of myofibers
Dear experts of histonet, Given my formal background (chemistry synthesis) and current experience (cardiovascular research) my question may seem either too simple of not well directed. Please forgive my inexperience in these matters. In short, I am looking for a simple method that enhances the fluorescence of the cardio myofibers. In our institution, we have constructed a special setup allowing for investigation of vessel morphology. This is carried out by infusing a fluorescent plastic that polymerizes into an organ and then serially slicing the specimen while after each slice a high resolution image is taken of the remaining bulk material. As such, we can create high resolution 3D images of the vasculature, e.g. of a heart or kidney. Recently we have extended our setup (we are using a 16mpixel cooled camera which also allows very long exposure times) and are now trying to visualize the muscle fibers of the heart. For this we use a powerled (around 400nm) and image at around 600nm which seems to generates images that show autofluorescence of either collagen or muscle (I am unsure which this may be). Given these initial promising results, I would like to visualize the muscle fibers in more detail by using a staining protocol that would allow to stain post mortem hearts. I have gathered information so far that includes the use of ALA or Bouin's solution but there may be much better protocols suitable for this problem. I appreciate all feedback, thank you in advance, Jeroen Jeroen PHM van den Wijngaard, PhD Department of Biomedical Engineering and Physics Academic Medical Center Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands Tel: +31 (20) 5668796 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] von kossa
I did a von kossa day the other since it was sunny out and never thought to use a hema counterstain. I will try that next time! I use ammonia water too as a bluing agent. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of histonet-requ...@lists.utsouthwestern.edu Sent: Friday, July 17, 2009 1:07 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 68, Issue 21 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. Hi Jan, (Carrie Disbrow) 2. Quality Stuff (kristen arvidson) 3. RE: Quality Stuff (Mike Pence) 4. Re: Quality Stuff (Rene J Buesa) 5. Re: Biological hood with grossing station (Joseph Saby) 6. RE: Hi Jan, (Tony Henwood) 7. Re: Hi Jan, (Victor Tobias) 8. Re: Quality Stuff (theci...@yahoo.com) 9. VonKossa's calcium stain (karine cadoret) 10. RE: VonKossa's calcium stain (Tony Henwood) 11. Re: VonKossa's calcium stain (Jack Ratliff) 12. Protocol for fluorescence of myofibers (J.P.H.M. van den Wijngaard) 13. NSH Meeting in Alabama (mtitf...@aol.com) -- Message: 1 Date: Thu, 16 Jul 2009 16:15:24 -0400 From: Carrie Disbrow dis...@shands.ufl.edu Subject: [Histonet] Hi Jan, To: histonet@lists.utsouthwestern.edu Message-ID: 4a5f521b.72ac.005...@shands.ufl.edu Content-Type: text/plain; charset=US-ASCII Hi Jan, Thanks for your input! I have two A. S. degrees. One in veterinary nursing/technology and one in histology. And I have an AA where all my electives were biology,chemistry and micro. Then I'll have a BS in veterinary nursing/management. I'm starting a molecular program in January. So, I have a strong skills in courses you mentioned. I'm sure I want to do the R D techniques but not so sure about the management! The other thing about a histology career is learning how vast the field is. Did anyone ever have a counselor in their program that explained the different types of positions? I'm looking forward to attending the NSH convention in October. It will be my first one! Carrie -- Message: 2 Date: Thu, 16 Jul 2009 14:14:36 -0700 (PDT) From: kristen arvidson arvidsonkris...@yahoo.com Subject: [Histonet] Quality Stuff To: histonet histonet@lists.utsouthwestern.edu Message-ID: 685857.31127...@web65709.mail.ac4.yahoo.com Content-Type: text/plain; charset=iso-8859-1 Hello, I work in a derm lab and we do all the grossing. We hand write on all of our blocks and slides, so you can imagine we have mislabelings from time-to-time. I was wondering if other labs have acceptable limits set for errors such as these, and if so what are they like? I am working on setting standards and corrective actions for errors in the lab. Thank you for any input. -- Message: 3 Date: Thu, 16 Jul 2009 16:21:00 -0500 From: Mike Pence mpe...@grhs.net Subject: RE: [Histonet] Quality Stuff To: kristen arvidson arvidsonkris...@yahoo.com, histonet histonet@lists.utsouthwestern.edu Message-ID: 661949901a768e4f9cc16d8af8f2838c017a3...@is-e2k3.grhs.net Content-Type: text/plain; charset=iso-8859-1 There is NO margin of error acceptable in mislabeling blocks or slides. I expect 100% compliance with this in my department. When you have like specimens all day like derm, you cannot make labeling errors. Mike -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of kristen arvidson Sent: Thursday, July 16, 2009 4:15 PM To: histonet Subject: [Histonet] Quality Stuff Hello, I work in a derm lab and we do all the grossing. We hand write on all of our blocks and slides, so you can imagine we have mislabelings from time-to-time. I was wondering if other labs have acceptable limits set for errors such as these, and if so what are they like? I am working on setting standards and corrective actions for errors in the lab. Thank you for any input. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Message: 4 Date: Thu, 16 Jul 2009 14:47:23 -0700 (PDT) From: Rene J Buesa rjbu...@yahoo.com Subject: Re: [Histonet] Quality Stuff To: histonet histonet@lists.utsouthwestern.edu,
[Histonet] RE: Histonet Digest, Vol 68, Issue 21
Why dont you use NFR as counterstain for Von kassa? From: histonet-requ...@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 68, Issue 21 To: histonet@lists.utsouthwestern.edu Date: Fri, 17 Jul 2009 10:02:55 -0700 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. Hi Jan, (Carrie Disbrow) 2. Quality Stuff (kristen arvidson) 3. RE: Quality Stuff (Mike Pence) 4. Re: Quality Stuff (Rene J Buesa) 5. Re: Biological hood with grossing station (Joseph Saby) 6. RE: Hi Jan, (Tony Henwood) 7. Re: Hi Jan, (Victor Tobias) 8. Re: Quality Stuff (theci...@yahoo.com) 9. VonKossa's calcium stain (karine cadoret) 10. RE: VonKossa's calcium stain (Tony Henwood) 11. Re: VonKossa's calcium stain (Jack Ratliff) 12. Protocol for fluorescence of myofibers (J.P.H.M. van den Wijngaard) 13. NSH Meeting in Alabama (mtitf...@aol.com) -- Message: 1 Date: Thu, 16 Jul 2009 16:15:24 -0400 From: Carrie Disbrow dis...@shands.ufl.edu Subject: [Histonet] Hi Jan, To: histonet@lists.utsouthwestern.edu Message-ID: 4a5f521b.72ac.005...@shands.ufl.edu Content-Type: text/plain; charset=US-ASCII Hi Jan, Thanks for your input! I have two A. S. degrees. One in veterinary nursing/technology and one in histology. And I have an AA where all my electives were biology,chemistry and micro. Then I'll have a BS in veterinary nursing/management. I'm starting a molecular program in January. So, I have a strong skills in courses you mentioned. I'm sure I want to do the R D techniques but not so sure about the management! The other thing about a histology career is learning how vast the field is. Did anyone ever have a counselor in their program that explained the different types of positions? I'm looking forward to attending the NSH convention in October. It will be my first one! Carrie -- Message: 2 Date: Thu, 16 Jul 2009 14:14:36 -0700 (PDT) From: kristen arvidson arvidsonkris...@yahoo.com Subject: [Histonet] Quality Stuff To: histonet histonet@lists.utsouthwestern.edu Message-ID: 685857.31127...@web65709.mail.ac4.yahoo.com Content-Type: text/plain; charset=iso-8859-1 Hello, I work in a derm lab and we do all the grossing. We hand write on all of our blocks and slides, so you can imagine we have mislabelings from time-to-time. I was wondering if other labs have acceptable limits set for errors such as these, and if so what are they like? I am working on setting standards and corrective actions for errors in the lab. Thank you for any input. -- Message: 3 Date: Thu, 16 Jul 2009 16:21:00 -0500 From: Mike Pence mpe...@grhs.net Subject: RE: [Histonet] Quality Stuff To: kristen arvidson arvidsonkris...@yahoo.com, histonet histonet@lists.utsouthwestern.edu Message-ID: 661949901a768e4f9cc16d8af8f2838c017a3...@is-e2k3.grhs.net Content-Type: text/plain; charset=iso-8859-1 There is NO margin of error acceptable in mislabeling blocks or slides. I expect 100% compliance with this in my department. When you have like specimens all day like derm, you cannot make labeling errors. Mike -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of kristen arvidson Sent: Thursday, July 16, 2009 4:15 PM To: histonet Subject: [Histonet] Quality Stuff Hello, I work in a derm lab and we do all the grossing. We hand write on all of our blocks and slides, so you can imagine we have mislabelings from time-to-time. I was wondering if other labs have acceptable limits set for errors such as these, and if so what are they like? I am working on setting standards and corrective actions for errors in the lab. Thank you for any input. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Message: 4 Date: Thu, 16 Jul 2009 14:47:23 -0700 (PDT) From: Rene J Buesa rjbu...@yahoo.com Subject: Re: [Histonet] Quality Stuff To: histonet histonet@lists.utsouthwestern.edu, kristen arvidson arvidsonkris...@yahoo.com Message-ID: 720913.73313...@web65707.mail.ac4.yahoo.com Content-Type: text/plain; charset=utf-8 There are no acceptable standards for mistakes. The present tendency of implementing the 6σ
Re: [Histonet] NSH Meeting in Alabama
I agree Michael!!! Phyllis Thaxton HT(ASCP)QIHC DCH Regional Medical Center Tuscaloosa, AL From: mtitf...@aol.com mtitf...@aol.com To: histonet@lists.utsouthwestern.edu Sent: Friday, July 17, 2009 10:33:34 AM Subject: [Histonet] NSH Meeting in Alabama With the NSH Annual Convention in Alabama this year, I don't want any corny jokes on the Histonet?about my adopted home state!? Might hurt my feelings! Michael Titford Pathology USA Mobile AL ___ 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] What percent of HTL's do not have a BS degree?
Here, Here! Try getting a job with a straight BS anyway... I had to go back to school to get a marketable job and was going to go for an MLT degree until I saw the Histology program in the tech school catalogue. This is what I have always wanted to do without knowing the name for it. Besides, the phlebotomy bit makes me a bit squeemish. I can deal with anything in the gross room or morgue with no problem though. :) I love the lifetime of learning bit, and so far I have been lucky to be able to indulge my curiosity and learn more. I essentially have 3 degrees (BS Biology w/ a major in English, and an AA in Histotechnology) for my HTL, which I took because who knows what the future may hold. Besides, why pay to take both exams? Not interested in managerial stuff though, just like to rat around in the lab. I have to order supplies and keep up the paperwork. That is plenty for me thanks. I like to get my hands dirty. (darn Schiffs) I have actually known a few people who have worked as histologists while waiting to get into med school. Doesn't sound dead-end to me. There are many ways histology can be used as a starting place for other jobs with more ceiling space. Creativity and ambition are the keys. (and a bit more hard work) Nothing worthwhile is ever easy. Claire P.S. Anyone about to nit-pic my grammar, I have a English Major with an emphasis in Literature. Never could diagram sentences. From: histonet-boun...@lists.utsouthwestern.edu on behalf of Wahlberg, Nikki Sent: Tue 7/14/2009 6:02 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] What percent of HTL's do not have a BS degree? I would just like to add that in my opinion it is people who make statements like the one below that are holding our field back from being seen as a career. The hospitals as well as the doctors are also to blame. I am very proud to have a B.S. and A.S.S. degree and an HTL certification. I would really like to see a monkey do my job and still achieve the high GLP standards and high quality of work that is required to get medical devices approved for human use. It makes me sad to hear people say that this is just a job not a career. I do not believe that anyone should be allowed to just come off the street and do our job. It up to us as a community to demand that institutions require certification and recognize our educations. I don't know about anyone else out there but my education cost me a lot of money and will keep me in debt for many years. I didn't waste all that money on just a job this is my career and I am very proud of the work I do. Nikki ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] nuclear bubbling
How about microwaving to dry slides? Can that cause nuclear bubbling? When you say completely drained off, does that mean your slides have to be completely dry prior to placing them in the oven? Thanks, Sandy -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Wednesday, July 15, 2009 2:59 PM To: Histonet; Joyce Cline Subject: Re: [Histonet] nuclear bubbling Anybody can experience nuclear bubbling in any type tissue as long as the sections as set to dry at high temperature BEFORE they are completely drained off! René J. --- On Wed, 7/15/09, Joyce Cline jcl...@wchsys.org wrote: From: Joyce Cline jcl...@wchsys.org Subject: [Histonet] nuclear bubbling To: Histonet histonet@lists.utsouthwestern.edu Date: Wednesday, July 15, 2009, 3:45 PM Has anyone experienced nuclear bubbling on prostate biopsies? Joyce * CONFIDENTIALITY NOTICE * 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. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Fat histology
I work with bone and connective tissue. I have just been asked to process and section some fat. I would like to get some information on how to work with fat. Any information will be very helpful. -- */-- Karen Bowden Staff Research Associate II University of CA, San Diego Department of Orthopedics 9500 Gilman Dr. 0630 La Jolla, CA 92093-0630 858-534-4655 voice 858-534-5304 fax CONFIDENTIALITY NOTICE: THE INFORMATION TRANSMITTED IN THIS E-MAIL IS INTENDED ONLY FOR THE PERSON OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN CONFIDENTIAL AND/OR PRIVILEGED MATERIAL. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR OTHER USE OF OR TAKING OF ANY ACTION IN RELIANCE UPON THIS INFORMATION BY PERSONS OR ENTITIES OTHER THAN THE INTENDED RECIPIENT IS PROHIBITED. IF YOU RECEIVED THIS E-MAIL IN ERROR, PLEASE CONTACT THE SENDER AND DELETE THE MATERIAL FROM ANY COMPUTER./* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] NSH Meeting in Alabama
Bless your li'l ol' hearts y'all, we wouldn't dream of it! However, I will accustom myself to grits and okra before leaving for Birmingham! Tim Morken UCSF Medical Center San Francisco, CA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Phyllis Thaxton Sent: Friday, July 17, 2009 12:08 PM To: mtitf...@aol.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] NSH Meeting in Alabama I agree Michael!!! Phyllis Thaxton HT(ASCP)QIHC DCH Regional Medical Center Tuscaloosa, AL From: mtitf...@aol.com mtitf...@aol.com To: histonet@lists.utsouthwestern.edu Sent: Friday, July 17, 2009 10:33:34 AM Subject: [Histonet] NSH Meeting in Alabama With the NSH Annual Convention in Alabama this year, I don't want any corny jokes on the Histonet?about my adopted home state!? Might hurt my feelings! Michael Titford Pathology USA Mobile AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Fat histology
Hi Karen, I had the same problem recently. I got very good advise trough the histonet for working with fat. You might want to check the arhives. In my case, I was in a big time crunch with this dog fat, so after all the fat was collected and stored for a while in 10% NBF I did'nt have any time to post fix it, I just had to put it in the processor. It came out great! I used 1 hour in all steps and the steps are: 1Formalin1 hour 280% 1 hour 395% 1 hour 495% 1 hour 5100%1 hour 6100%1 hour 7100 % 1 hour 8Clear Rite 1 hour 9Clear Rite 1 hour 10 Clear Rite 1 hour 11 Paraffin 1 hour 12 Paraffin 1 hour 13 Paraffin 1 hour 14 Paraffin 1 hour I did it again few days ago (the same way) and again had great result. We also did IF on this fat and it worked great as well. Good luck with it Michelle Research Specialist USC Keck School of Medicine Los Angeles, CA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] VonKossa's calcium stain. Counterstain etc
Any haemalum will do, but blue is not the most pleasing contrast to go with the black silver deposits. Pink or light red is better. Here are three pink to red counterstains, all traditional. Check in a textbook or manual for instructions. 1. Neutral red (CI 50040) is good: 0.5% in water; adjust to pH4 with acetic acid; stain for about 2 minutes; the solution keeps for at least 5 years and can be used repeatedly. 2. Safranine O (CI 50240) can be used similarly but needs a longer staining time. 3. Nuclear fast red (CI 60760) is also OK: NFR 0.2G, aluminium sulphate crystals 10G, water 200ml; heat until it boils, cool overnight, decant and filter; stain for 5-10min. The solution is good for about a year; always filter before using. The Biological Stain Commission has standards for certification of all three of these dyes. Certified neutral red and safranine O have been available for many (50+) years. Nuclear fast red was only recently added to the BSC's list. See Frank et al 2007. Certification procedures for nuclear fast red (Kernechtrot), C.I. 60760. Biotechnic Histochemistry 82: 35-39. Certified NFR powder may not yet be available to labs or to vendors of stain solutions. If you buy a ready-made solution of any dye you should choose one that was made from a BSC-certified batch of the powder. To any vendors of dye powders who read this message: email me for more information about the certification criteria for nuclear fast red. Also, check out http://www.biologicalstaincommission.org and click on one of the Vendors tabs or links. John Kiernan Anatomy, UWO London, Canada = = = - Original Message - From: karine cadoret kcado...@amc.edu.au Date: Thursday, July 16, 2009 22:36 Subject: [Histonet] VonKossa's calcium stain To: histonet@lists.utsouthwestern.edu Hi, When doing a VonKossa stain in order to demonstrate calcium in tissue, does it matter much if I use Mayer's hematoxylin instead of Ehrlich's hematoxylin (which takes 6 months to ripen) ? Also, can I simply use homemade scott's tapwater for blueing instead of using a lithium carbonate solution ? Thank you for your help, Karine Cadoret Fish health laboratory manager National Center for Marine Conservation and Resource Sustainability Newnham, TAS Australia ___ 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
