[Histonet] Frozen section ...HELP
Hi all, after more than a decade of NOT cutting frozen sections, I find myself back at the ice-face. To get my hand in (not literally) I thought I would do some trial sections on stored tissue - stuff that was in formalin and now in 70% alcohol. Horror ; dismay. The tissue, once frozen, is all mushy in the middle. Is this because of teh long storage? is there anything I can do to improve the situation? much appreciated -- 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
Re: [Histonet] Frozen section ...HELP
The freezing point of water is 0 degrees C. The freezing point of 100% ethanol is -114 degrees C. The freezing point of 70% alcohol is about -48 degrees C. Since most cryostats are at -20 to -25 degrees C, your tissue isn't freezing completely. You probably have slush ice inside the cells. Not hard enough to support the tissue during a frozen section. Fresh tissue would be the best, as formalin fixed tissue also tends to cut awful (whole different reason). If you can't get fresh tissue from human or animal necropsy, can you get some parts from a raw uncooked chicken to practice on? Save some muscle, skin, liver, etc., from tonight's supper? Peggy A. Wenk, HTL(ASCP)SLS Beaumont Hospital Royal Oak, MI 48073 -- From: louise renton louise.ren...@gmail.com Sent: Monday, November 08, 2010 4:43 AM To: Histonet Histonet@lists.utsouthwestern.edu Subject: [Histonet] Frozen section ...HELP Hi all, after more than a decade of NOT cutting frozen sections, I find myself back at the ice-face. To get my hand in (not literally) I thought I would do some trial sections on stored tissue - stuff that was in formalin and now in 70% alcohol. Horror ; dismay. The tissue, once frozen, is all mushy in the middle. Is this because of teh long storage? is there anything I can do to improve the situation? much appreciated -- 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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] shrinkage during IHC
In my experience, IHC usually plumps tissue back up during reteival. Is this noticed with every antibody or just a few? Certain tissue types? -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of zodia...@comcast.net Sent: Saturday, November 06, 2010 8:28 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] shrinkage during IHC To All, We are a lab that sends our specimens out for IHC and have just switched to another reference laboratory for these services. Our pathologist is saying that the tissue looks shrunk on the IHC slides, yet the slides that I process (HE, and special stains)are fine. Does anyone know what is causing this? The reference lab said it could be the type of slides that I use to mount the sections we send to them. My knowledge in IHC is limited. Also, if this helps, they are FFPE tissue. Thanks for your help Jenny ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information in this e-mail, and any attachments therein, is confidential and for use by the intended addressee only. If this message is received by you in error please do not disseminate or read further. Please reply to the sender that you have received the message in error, then delete the message. Although Catholic Health Services of Long Island attempts to sweep e-mail and attachments for viruses, it does not guarantee that either are virus-free and accepts no liability for any damage sustained as a result of viruses. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Frozen section ...HELP
Hopefully you removed the antifreeze (alcohol) before freezing? :-) I have run into this a few times. I work in a core facility where people send me samples from all over. Recently someone sent me some samples fixed in Histofix, which is a commercially available aqueous fixative. But they neglected to tell me they add 15% ethanol to the commercial solution. I froze all the samples in OCT compound, but they could not be sectioned. I had to melt them all, soak them in several changes of buffer for several hours to remove the OCT and the methanol, then put them in fresh OCT and refreeze them. And that was only 15% alcohol, not 70%! ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Distance Learning
We are thinking of using one of the online or distance learning programs to help in the training of new histotechs. Has anyone had any experience with any of these programs, what colleges offer the program, pros, cons. Any feedback is welcome. Thanks Phyllis Thaxton HT(ASCP)QIHC DCH Regional Medical Center Tuscaloosa, AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Microwave processing effect on DNA/RNA
Hello All, I have seen some posts on the possible damaging effects on DNA and RNA by microwave processing. Does anyone have a reference article that they can share with me? We are in the process of budgeting for next year and we are looking at microwave processors; but I want to verify if this claim is or is not valid. Thank you, Matt Brooks, BS, HT (ASCP) Histology Supervisor InCyte Pathology mbro...@incytepathology.com 509-892-2744 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: 8. Distance Learning (Phyllis Thaxton)
Hi Everyone, I'm new to histonet and haven't participated in any discussions yet, but I noticed the question regarding distance/online programs for histotechnicians and I'm currently enrolled in an accredited Histology Technician program through Columbus State Community College. I'd be more than happy to share any feedback as a student if you're interested. Feel free to email me privately at: vkline2...@comcast.net -Victoria Kline ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Frozen Section Help
Did you cryoprotect the tissue before you cut it? JB Jo Ann Biedermann Research Assistant University of Arkansas for Medical Sciences Reynolds Institute on Aging 629 Jack Stephens Drive Room 3173 Mail Slot 807 Little Rock, AR 72205 Phone: 501-526-5803 FAX: 501-526-5830 jabiederm...@uams.edu -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of histonet-requ...@lists.utsouthwestern.edu Sent: Monday, November 08, 2010 12:06 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 84, Issue 9 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. cytotechnology (Mimmo Pozzuoli) 2. RE: SPAM-LOW: [Histonet] shrinkage during IHC (Patsy Ruegg) 3. RE: SPAM-LOW: [Histonet] cytotechnology (Patsy Ruegg) 4. Frozen section ...HELP (louise renton) 5. Re: Frozen section ...HELP (Lee Peggy Wenk) 6. RE: shrinkage during IHC (Kuhnla, Melissa) 7. RE: Frozen section ...HELP (Monfils, Paul) 8. Distance Learning (Phyllis Thaxton) 9. Microwave processing effect on DNA/RNA (Matt Brooks) -- Message: 1 Date: Sun, 7 Nov 2010 13:36:09 -0500 From: Mimmo Pozzuoli mimmopozzu...@gmail.com Subject: [Histonet] cytotechnology To: histonet@lists.utsouthwestern.edu Message-ID: aanlktim4e0d0+lofa=wtkcprr+thep7h3ew7-b=tk...@mail.gmail.com Content-Type: text/plain; charset=ISO-8859-1 Are any histotechs out there performing FISH or automated cyto procedures in their labs? If so, are your pathologists having to screen and act as the cyto supervisors? What are the guidelines on this? Can a histotech perform cytoprep functions, or is the slide screening integral to the position? -- Message: 2 Date: Sun, 7 Nov 2010 13:20:58 -0700 From: Patsy Ruegg pru...@ihctech.net Subject: RE: SPAM-LOW: [Histonet] shrinkage during IHC To: zodia...@comcast.net, histonet@lists.utsouthwestern.edu Message-ID: 9109304a049d40f889c150360425e...@prueggihctechlt Content-Type: text/plain; charset=us-ascii You should have the same morphology from the IHC slides done outside that you get from your HE, do you provide them with slides or the Block? The only thing I can think of is if they airdry after IHC instead of going thru alcohols and xylene to coverslip (if they use AEC or ap/red they may airdry), I have seen cell shrinkage from airdrying after IHC occasionally. Something else just occurred to me, if they over heat during HIER I suppose this could happen, but I have not seen it. Regards, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. Ste.215 Aurora, CO 80045 720-859-4060 fax 720-859-4110 www.ihctech.net www.ihcrg.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of zodia...@comcast.net Sent: Saturday, November 06, 2010 6:28 AM To: histonet@lists.utsouthwestern.edu Subject: SPAM-LOW: [Histonet] shrinkage during IHC To All, We are a lab that sends our specimens out for IHC and have just switched to another reference laboratory for these services. Our pathologist is saying that the tissue looks shrunk on the IHC slides, yet the slides that I process (HE, and special stains)are fine. Does anyone know what is causing this? The reference lab said it could be the type of slides that I use to mount the sections we send to them. My knowledge in IHC is limited. Also, if this helps, they are FFPE tissue. Thanks for your help Jenny ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Message: 3 Date: Sun, 7 Nov 2010 13:55:42 -0700 From: Patsy Ruegg pru...@ihctech.net Subject: RE: SPAM-LOW: [Histonet] cytotechnology To: 'Mimmo Pozzuoli' mimmopozzu...@gmail.com, histonet@lists.utsouthwestern.edu Message-ID: fcfb1eba62d341d29251230913eb0...@prueggihctechlt Content-Type:
[Histonet] Re: Microwave processing effect on DNA/RNA
Matt, You asked an interesting question and it got me to googling. Since microwave radiation is non-ionizing it should not adversely affect things like DNA and RNA. I found this summary of a group of publications on this and apparently in 1995, Kakita demonstrated that microwaves were capable of fragmenting viral DNA, and they were able to conclude it was not due to the heating effect. http://www.rfsafe.com/research/rf_hazards/dna_damage/microwave_effect.htm Most of the data I can find seems to correspond to the type of electromagnetic radiation produced by cell phones, because that's the biggie going around these days. Here's a more recent paper on cell phone microwaves and their effect on human lymphocytes. It might be useful to check out the reference list, especially the ones cited in the introduction. http://www.hese-project.org/hese-uk/en/papers/sarimov_chromatin_heatshock_ieee04.pdf The thing I don't know is how the microwaves from cell phones equate, or if they even do, with microwave oven exposure, especially since we can control the samples from being overheated. Also, this study was done on cell cultures, which while it might give interesting and useful information, it does not mimic the conditions of a whole organism. In addition, the microwave effects were reported on unfixed cells, unlike the samples you will be processing. What I would do if I were you is get a demo unit in, simulaneously process 10-20 different patient samples with the microwave processor and with your routine processor after your routine fixation, and then send them off for analysis. That should tell you all you need to know right there. If anyone has already done this, please speak up! Best wishes, Teri Johnson, HT(ASCP)QIHC Managing Director, Histology Facility Stowers Institute for Medical Research Kansas City, MO ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] p53 cytology controls
We receive requests from our pathologists for p53 IHC on urine but I am unsure as to what controls to use. We currently use a FFPE tissue control (I understand that it is not correct) but up until now this issue has not been addressed. Can anyone help with good cytology controls? Thanks. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] p53 cytology controls
Hi Justin, Do you run the p53 on a cell block or a cytospin or something different? Mark On Mon, Nov 8, 2010 at 12:36 PM, Justin Peters jpet...@bostwicklaboratories.com wrote: We receive requests from our pathologists for p53 IHC on urine but I am unsure as to what controls to use. We currently use a FFPE tissue control (I understand that it is not correct) but up until now this issue has not been addressed. Can anyone help with good cytology controls? Thanks. ___ 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
