[Histonet] QC on stained slides
Hi all As part of a self assessment programme conducted by my employer, and related to my performance review and salary adjustment, I need to determine the criteria of what makes a stained slide acceptable or unacceptable. I was wondering if anyone out there had a checklist that they would be willing to share, that i could perhaps adapt. I realise that the easiest would be to send slides out for external control, but in this case it is not feasible. What I put together is this: - Quality of decalcification, processing, infiltration - Quality of sections (no wrinkles, missing bits, scores etc) - Entire representation of tissue area - staining pattern as expected according to protocol - coverslipped without bubbles or other inclusions - labelled neatly and correctly but, the question inmy mind is what would be the criteria that would make a slide merely adequate or truely outstanding? PLease help thank you -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa +27 11 717 2298 (tel fax) 073 5574456 (emergencies only) There are nights when the wolves are silent and only the moon howls. George Carlin No trees were killed in the sending of this message. However, many electrons were terribly inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] decalcification
I would appreciate any feedback on what all are using in your decalcification process. We get a lot of large bones in and the past 2-3 months have noticed a huge problem in our microtomy process with these samples. We have been grossing the bones in and leaving the sample in the cassette in 10% formalin for 24 hours befoere placing in decal for up to 8 hours and still having the inner portion of the sample look underprocessed and crunchy! Any suggestions would be appreciated! Dorothy Webb, HT Regions Hospital Technical Supervisor 651-254-2962 This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] decalcification
I have used formical for years and it works great. You don'= t have
to pre-fix your samples because there is formalin in with the decal.
It fixes and decals at the same time. I don't know how many sp ecimens
you have, but a sure fire way to check is to weigh the bone
after g= rossing. Then everyday weigh it and replace the decal
solution. = It should start losing weight because the calcium is
coming out. As = soon as the sample starts gaining weight, take it
out. Now it is just= absorbing fluid. In the past when I have done
huge bones, I use this= technique (learned at a NSH convention) and it
works like a charm! H= ope that helps =)
PS-The formical is made by Dec= al company.
Sarah Goebel, B.A., HT (ASCP)
Histotechnician
= /span
XBiotech USA Inc.
8201 East Riverside Dr. Bldg 4 = Suite 100
Austin, Texas 78744
(512)386-5107
Original Message
Subject: [Histonet] decalcification
From: Webb, Dorothy L [1]dorothy.l.w...@healthpartners.com
Date: Mon, August 02, 2010 7:14 am
To: '[2]histo...@lists.u= tsouthwestern.edu'
[3]histo...@lists.uts= outhwestern.edu
I would appreciate any feedback on what all are using in your
decalcificati= on process. We get a lot of large bones in and the past
2-3 months have no= ticed a huge problem in our microtomy process with
these samples. We have = been grossing the bones in and leaving the
sample in the cassette in 10% fo= rmalin for 24 hours befoere placing
in decal for up to 8 hours and still ha= ving the inner portion of the
sample look underprocessed and crunchy! Any = suggestions would be
appreciated!
Dorothy Webb, HT
Regions Hospital Technical Supervisor
651-254-2962
This e-mail and any files transmitted with it are confidential and are
inte= nded solely for the use of the individual or entity to whom they
are addres= sed. If you are not the intended recipient or the
individual responsible fo= r delivering the e-mail to the intended
recipient, please be advised that y= ou have received this e-mail in
error and that any use, dissemination, forw= arding, printing, or
copying of this e-mail is strictly prohibited.
If you have received this e-mail in error, please immediately notify
the He= althPartners Support Center by telephone at (952) 967-6600.
You will be rei= mbursed for reasonable costs incurred in notifying
us. HealthPartners R001.= 0
___
Histonet mailing list
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References
1. 3Dmailto:dorothy.l.w...@healthpartners 2.
3Dmailto:histonet@lists.utsouthwestern.edu;
3. 3Dmailto:histonet@lists.utsouthwestern.edu;
4. 3Dmailto:Histonet@lists.utsouthwestern.edu;
5. 3Dhttp://lists.utsouthwestern.edu/mailman/listinfo/histonet;
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Re: [Histonet] decalcification
You cannot set a fixed time for decalcification. You have to leave the bone in the decalcifying solution until iy is soft and ready for processing. René J. --- On Mon, 8/2/10, Webb, Dorothy L dorothy.l.w...@healthpartners.com wrote: From: Webb, Dorothy L dorothy.l.w...@healthpartners.com Subject: [Histonet] decalcification To: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu Date: Monday, August 2, 2010, 10:14 AM I would appreciate any feedback on what all are using in your decalcification process. We get a lot of large bones in and the past 2-3 months have noticed a huge problem in our microtomy process with these samples. We have been grossing the bones in and leaving the sample in the cassette in 10% formalin for 24 hours befoere placing in decal for up to 8 hours and still having the inner portion of the sample look underprocessed and crunchy! Any suggestions would be appreciated! Dorothy Webb, HT Regions Hospital Technical Supervisor 651-254-2962 This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 ___ 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] Irving Ht Position
Nueterra, a leading developer and manager of physician-owned surgery centers and specialty hospitals has an immediate opportunity available for a Histotechnician. Great opportunity for a Histotechnician in a brand new laboratory! Nueterra Pathology Services in Irving, TX is looking for a certified HT or HTL to run their newly constructed laboratory. Candidate must be ASCP certified and CLIA certified to perform gross dissection, prior supervisory experience preferred. The candidate will be responsible for the following: Creation and maintenance of policies and procedures to CLIA standards, leading lab through CLIA inspection, maintenance and quality control for equipment, and routine histology duties. The position offers a competitive salary, medical/dental insurance, 401K plan, and vacation/sick leave. Interested applicants should contact Meredith Hale phone 214-596-2219 or through email at mh...@carisdx.com mailto:mh...@carisdx.com . Meredith Hale HT (ASCP) Operations Liaison Director and Education Coordinator Caris Life Sciences 6655 North MacArthur Blvd, Irving Texas 75039 direct: 214-596-2219 cell: 469-648-8253 fax: 972-929-9966 mh...@carisls.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] decalcification
Hi Dorothy, I'm not sure what size bone specimens with which you are working, but I generally work with femurs calvaria from rabbits, canine other small rodents. I fix my specimens longer in 10% NBF using an automated vacuum infiltration processor. Following fixation, if decal is required, I will decal in buffered formic acid solution and use a calcium reagent set to titrate to the decalcification endpoint. If you would like any specifics, contact me offline. Best regards, ~Sean McBride Senior Researcher Bone Tissue Engineering Center Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 412-268-8275 (o) 412-915-1683 (m) 412-268-9807 (fax) smcbr...@andrew.cmu.edu --- On Mon, 8/2/10, Webb, Dorothy L dorothy.l.w...@healthpartners.com wrote: From: Webb, Dorothy L dorothy.l.w...@healthpartners.com Subject: [Histonet] decalcification To: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu Date: Monday, August 2, 2010, 10:14 AM I would appreciate any feedback on what all are using in your decalcification process. We get a lot of large bones in and the past 2-3 months have noticed a huge problem in our microtomy process with these samples. We have been grossing the bones in and leaving the sample in the cassette in 10% formalin for 24 hours befoere placing in decal for up to 8 hours and still having the inner portion of the sample look underprocessed and crunchy! Any suggestions would be appreciated! Dorothy Webb, HT Regions Hospital Technical Supervisor 651-254-2962 This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 ___ 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] Collagen Type II antibody
I am in need of a good collagen Type II antibody that could be used in both rat and human samples. We would prefer a polyclonal antibody but might settle for a monoclonal. Anyone have any good suggestions? We will be staining the samples with DAB. Thanks in advance. Joel Reichensperger -- Joel Reichensperger Researcher II Southern Illinois University Plastic Surgery Institute jreichensper...@siumed.edu 217-545-7309 (Office) 217-545-1824 (Fax) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Collagen Type II antibody
Joel We use an antibody we purchase from MD Biosciences - its quite expensive but we find it works really well for us on multiple species with chondroitinase digestion. Protocol: Rabbit anti Human Collagen Type II Clone: Polyclonal Vendor: MD Biosciences Catalog Number: MD20211 Species: Rabbit Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Joel Reichensperger Sent: Monday, August 02, 2010 11:30 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Collagen Type II antibody I am in need of a good collagen Type II antibody that could be used in both rat and human samples. We would prefer a polyclonal antibody but might settle for a monoclonal. Anyone have any good suggestions? We will be staining the samples with DAB. Thanks in advance. Joel Reichensperger -- Joel Reichensperger Researcher II Southern Illinois University Plastic Surgery Institute jreichensper...@siumed.edu 217-545-7309 (Office) 217-545-1824 (Fax) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] RE: Histonet Digest, Vol 81, Issue 1
Jennifer, I don't use the Mach 1 for animal tissue. I am working with rat tissue and use the Biocare Rabbit on Rodent or Mouse on Rat HRP or AP Polymers. I have absolutely no background and my sections are 40um thick. Perhaps you are using the wrong kit. Hope this helps, Tina -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Perry, Margaret Sent: Monday, August 02, 2010 10:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Histonet Digest, Vol 81, Issue 1 What antibody are you trying to use? -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of histonet-requ...@lists.utsouthwestern.edu Sent: Sunday, August 01, 2010 12:01 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 81, Issue 1 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. Help with Biocare polymer detection kit!! (Jennifer Campbell) -- Message: 1 Date: Sat, 31 Jul 2010 12:03:39 -0700 From: Jennifer Campbell jcampb...@vdxpathology.com Subject: [Histonet] Help with Biocare polymer detection kit!! To: histonet@lists.utsouthwestern.edu Message-ID: 5658cbdb9eae6545abe50d2563d81bf8181...@vdxserver01.vdxpathology.local Content-Type: text/plain; charset=iso-8859-1 I have been trying to get the Mach 1 polymer kit from Biocare to work on my K9 and feline tissue and I am getting terrible nonspecific staining in my positive and negative controls. It appears to be sticking to something in the glands of the intestinal epithelia. I am also getting nonspecific staining in my LN tissue. I decided to switch over to polymer detection kit because I had been dealing with the issue of endogenous biotin when using the streptavidin biotin-based detection system, and now I am getting this other nonspecific staining! I have tried doing runs eliminating my protein block and eliminating the negative control and primary antibody step altogether (to see if maybe the protein block or the diluent used in my negative control and antibody dilutions are causing the problem). I have also reduced the incubation time of both the probe and polymer from 15 min and 30 min, respectively, to 10 min each. I have tried cutting down my antigen retrieval time substantially as well. I'm not sure what else could be causing this. Any suggestions?? Thanks in advance, Jennifer Campbell -- ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 81, Issue 1 *** ___ 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] Beetles Head sectioning
Hi all: Does anyone has the proper procedures to obtain cryostat and/or paraffin sections of beetles head?. I have not been succesful at obtaining one. My samples cut well but during collection from the cryotome it tends to curl so I tend to loose the brain tissue. Additionally, is there any place that I can send my beetles head samples to be sectioned? This is to do basic brain measurements. I would really appreciate all possible feedback and pointing me to the right references if possible. Thanks Jorge L. Hurtado ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] (no subject)
hi there i need to know in a detail a special stain to demonstrate epilepsy in rat brain thanks a lot ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] decalcification
Dorothy- If your bones are indeed large, 24 hours in 10% NBF will not be sufficient for fixation. If the fixation isn't sufficient, none of the processes following that will be as effective as they might be. Even after trimming, I would recommend fixing an additional 24-48 hours for every mm of thickness of your bone section, at a minimum. If you are in a hospital setting and need reults quickly, you can use a processor (with heat, pressure and vacuum) to speed up the fixation. I use buffered formic acid to decal. The final concentration of formic acid is about 20-25%, so it is rather aggressive, but it also allows you to read the bone marrow after decal. If you need results in a short time (hospital setting again) you can use an aggressive reagent like RDO for decal, but keep in mind that nuclear detail will be lacking, but bone structure should still be readable. If you do not have the reagents to determine decal endpoints, you can (with training/practice) use a needle to pass through the sections to help determine if decal is complete. The needle should move through the specimen smoothly and not get hung up and stiff in the middle of the section. Flexibility of the bone section can tell you when you are appoaching the time when the needle test will give you useable results. Please do not read this as an endorsement for RDO or any other super aggressive decal solution. Personally, I always insist on being able to read all tissue elements, and these solutions, although fast, give less than optimal staining results. If you have further questions, please ask me off line. Joe Saby, BA HT From: Webb, Dorothy L dorothy.l.w...@healthpartners.com To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Sent: Mon, August 2, 2010 10:14:35 AM Subject: [Histonet] decalcification I would appreciate any feedback on what all are using in your decalcification process. We get a lot of large bones in and the past 2-3 months have noticed a huge problem in our microtomy process with these samples. We have been grossing the bones in and leaving the sample in the cassette in 10% formalin for 24 hours befoere placing in decal for up to 8 hours and still having the inner portion of the sample look underprocessed and crunchy! Any suggestions would be appreciated! Dorothy Webb, HT Regions Hospital Technical Supervisor 651-254-2962 This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 ___ 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] Yuba City HT Position
Histology Lab Tech. (Yuba City, CA) Licensed with a minimum of 3 years experience. Must be able to work with little supervision. Competitive salary and benefits. Please email resume' to rw...@northvalleygi.com for immediate consideration. * Location: Msvl/Yuba City * Compensation: DOE Meredith Hale HT (ASCP) Operations Liaison Director and Education Coordinator Caris Life Sciences 6655 North MacArthur Blvd, Irving Texas 75039 direct: 214-596-2219 cell: 469-648-8253 fax: 972-929-9966 mh...@carisls.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
