Re: [Histonet] Rolling of my ribbon ? Another paraffin question.
Just a thought - have you tried changing the angle of the blade? Each different type of blade from different vendors need a different angle for microtoming. Each different type of microtome from different vendors need a different angle for microtoming. I know these two facts, as I have tried different vendors' blades, and have had to change the angle on the microtome to get a good ribbon. The School also has different vendors' microtomes in the School, all using the same blades, and they had to be set at different angles. Plus, I have talked with vendors of various microtomes and various blades, and those who really know microtomy, especially those were were/are histotechs, will tell you that the blade angles need to be changed depending upon which blade is being used at which microtome. And also, from year to year, with different students sitting at the same microtome, we've also had to change the angles, depending on how the student cuts. We haven't changed brands of paraffin in the long time, but I'm wondering if we would need to change the angle of blade if we got a different paraffin. The easiest way to check is to move the knife angle all the way to one end (for example, on the vendor's microtome that we have the most of in the School, that would be to # 10.) Try microtoming. If that doesn't help, move it one degree (what would be #9 if there were a number). Try microtoming again. Keep doing this (changing angles and microtoming) until you get the best ribbon. (We found it easier to start at one end and move down, rather to start in the middle (say, #5), and then not know if we need to go up (#6) or go down (#4)). Let us know if knife angle is a factor with this new paraffin. I'd really like to know. Vendors - anyone know/have information that blade angle makes a difference with type of paraffin, for getting good ribbons? Peggy A. Wenk, HTL(ASCP)SLS -Original Message- From: Stella Mireles Sent: Wednesday, November 13, 2013 3:18 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Rolling of my ribbon ? Another paraffin question. *I am presently using a paraffin designated as an IM product.* *We are a facility that cuts only autopsies and have been experiencing alot of rolling of our sections.* *We did recently switch to this product, because of cost.* *Question : Is the paraffin you are using working well on autopsy tissue and producing ribbons right away ? Do you use it for infiltration as well as embedding ?* *Thank you for your assistant.* *Stella Walters* ___ 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] coverslipping question
Are you using tape or glass coverslips? We have found that when using tape coverslips, at least two very clean 100% alcohol steps must be added before slides go into the clearing reagent, be it xylene or Histoclear. If not, brown spots appear on some tissues, usually liver and bone. If there is any TRACE of water retained in the tissue, it will show up as a brown artifact on the tissue. This doesn't happen with conventional glass coverslipping and mounting medium - I think the mounting medium compensates for the retained moisture in the tissues. Jackie -Original Message- From: Kim Donadio one_angel_sec...@yahoo.com To: Gautier, Nicole M. nga...@lsuhsc.edu Cc: histonet histonet@lists.utsouthwestern.edu Sent: Wed, Nov 13, 2013 1:37 pm Subject: Re: [Histonet] coverslipping question I would try adding 1 or 2 more 100% alcohols before the histoclear. Sent from my iPhone On Nov 12, 2013, at 9:32 AM, Gautier, Nicole M. nga...@lsuhsc.edu wrote: My lab has been having a problem with specks appearing on our slides after they come out of Histoclear. Our protocol is to dehydrate in 2 minutes each of 70%, 80%, 95%, and 100% ethanol before 2 minutes 2 times in Histoclear. The slides are perfectly clear when they come out of the ethanols, but not when they come out of the Histoclear. Since it's not a problem we had when we first started, I tried changing out the Histoclear, but the problem remained. At the end of last week, I changed out everything - all glassware, all ethanols, all Histoclear. The problem went away and the 24 slides looked fine. Today, the slides are looking speckled again. So I guess my questions are: What could be causing the speckled appearance of the slides? and How often should I have to change the ethanols and Histoclear? I seem to remember years ago when I did this that we only changed the solutions monthly... Nicky Gautier Research Associate ___ 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
Re: [Histonet] neuropath stains
So far when I ask about neuro stains being automated by any company the answer is no. The volume clinically is not high enough and the times required would tie the stainers up from hours to overnight in for some steps. It would almost require a dedicated stainer for neuro only and would be very expensive. We do ours manually. Pam Marcum UAMS - Original Message - From: Felton Nails flna...@texaschildrens.org To: histonet@lists.utsouthwestern.edu Sent: Wednesday, November 13, 2013 4:46:04 PM Subject: [Histonet] neuropath stains Are the basic muscle and nerve biopsy stains being performed manually or has someone automated these stains? __ 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] (no subject)
I was just wondering if anyone was having difficulty cutting biopsies? We have been using the same blades for years and now it is so difficult to get a ribbon. There are three other area hospitals having the same problem. I have been trying samples from various vendors and the problem is still the same. Maybe the company that makes these microtome blades have cut costs and is now using a lower grade of metal for the blade. I don't know what it is, but it is driving everyone crazy. Also, we are spending more money on blades because they do not last as long and of course this does not make our Manager happy. I am just very frustrated. Any suggestions? LeAnn Murph, HT (ASCP) Aultman Hosptial Canton, Ohio Technical Specialist ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] (no subject)
I wouldn't be too quick to blame the blades. Have you changed paraffin brands/temperatures or changed any of your processing schedules? Is the fixative/fixation process the same? Is the collecting location doing anything different/have they had a change in staff? Also, what type of blades are you using? It would be easier for people to respond if they had this information. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Leann M. Murphy Sent: Thursday, November 14, 2013 11:04 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) I was just wondering if anyone was having difficulty cutting biopsies? We have been using the same blades for years and now it is so difficult to get a ribbon. There are three other area hospitals having the same problem. I have been trying samples from various vendors and the problem is still the same. Maybe the company that makes these microtome blades have cut costs and is now using a lower grade of metal for the blade. I don't know what it is, but it is driving everyone crazy. Also, we are spending more money on blades because they do not last as long and of course this does not make our Manager happy. I am just very frustrated. Any suggestions? LeAnn Murph, HT (ASCP) Aultman Hosptial Canton, Ohio Technical Specialist ___ 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: Embedding Paraffin
I use McCormick Paraplast Plus. On Tuesday, November 12, 2013 8:14 AM, Goins, Tresa tgo...@mt.gov wrote: Paraplast X-tra by McCormick. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Matthew D. Roark Sent: Tuesday, November 12, 2013 8:03 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Embedding Paraffin What paraffin does everyone like for embedding? We are currently using Surgipaths EM-400 but its dirty! Who has a clean, easy to section paraffin that they like? Thanks! Matthew Roark- HT/HTL(ASCP)CM Histology Specialist Saint Francis Medical Center 211 Saint Francis Drive Cape Girardeau, MO 63703 573-331-3982 mro...@sfmc.net http://www.sfmc.net/ ___ 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
Re: [Histonet] RE: Embedding Paraffin
We stumbled on Polyscientific's GemCut Pink Sapphire Parrafin and really like it. Before that we used Paraplast and it was OK but our blocks cut so much nicer with the GemCut. Andrea Grantham, HT (ASCP) Senior Research Specialist University of Arizona Cellular and Molecular Medicine Histology Service Laboratory P.O.Box 245044 Tucson, AZ 85724 algra...@email.arizona.edumailto:algra...@email.arizona.edu Tel: 520.626.4415 Fax: 520.626.2097 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Anyone using Section Transfer System by Microm?
Is anyone using the Thermo/Microm Secton Transfer System on their microtome? http://cellab.se/pdfbroschyr/Microm/STS.pdf I'd like to get some input for using it to cut serial sections of microarrays. Thanks! Tim Morken Supervisor, Electron Microscopy and Neuromuscular Special Studies UC San Francisco Medical Center Box 1656 505 Parnassus Ave San Francisco, CA 94143 USA 415.353.1266 (office) tim.mor...@ucsfmedctr.orgmailto:tim.mor...@ucsfmedctr.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] neuropath stains
Our immunofluorescence protein antibody stains are automated, but not the routine histochemical stains used for muscle biopsies. Between the importance of maintaining pH levels, incubation temperatures (SDH,NADH,COX, AtPase's, Amyloid, MAD, Myophos, Acid Phos) and the need of a rotator plate (ORO). Really the only options for automation would be the HE, Gomori Trichrome and PAS. Sarah E. Lewis HT, ASCP Laboratory Coordinator The Research Institute at Nationwide Children's Hospital Center for Gene Therapy Neuromuscular Division Rm WA3110 (614)722-2204 -Original Message- From: Nails, Felton [mailto:flna...@texaschildrens.org] Sent: Wednesday, November 13, 2013 5:46 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] neuropath stains Are the basic muscle and nerve biopsy stains being performed manually or has someone automated these stains? __ 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] RE: Anyone using Section Transfer System by Microm?
I would also like to know -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Morken, Timothy Sent: Thursday, November 14, 2013 9:55 AM To: Histonet Subject: [Histonet] Anyone using Section Transfer System by Microm? Is anyone using the Thermo/Microm Secton Transfer System on their microtome? http://cellab.se/pdfbroschyr/Microm/STS.pdf I'd like to get some input for using it to cut serial sections of microarrays. Thanks! Tim Morken Supervisor, Electron Microscopy and Neuromuscular Special Studies UC San Francisco Medical Center Box 1656 505 Parnassus Ave San Francisco, CA 94143 USA 415.353.1266 (office) tim.mor...@ucsfmedctr.orgmailto:tim.mor...@ucsfmedctr.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __ This message is confidential, intended only for the named recipient(s) and may contain information that is privileged or exempt from disclosure under applicable law. If you are not the intended recipient(s), you are notified that the dissemination, distribution, or copying of this message is strictly prohibited. If you receive this message in error, or are not the named recipient(s), please notify the sender at either the e-mail, fax, address, or telephone number listed above and delete this e-mail from your computer. Thank You. __ ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Has anyone had any problems with the Vector Immuedge pap pen?
Hi everyone, Has anyone had any problems with the Immedge Pap pen from Vector. Cat # H-4000 It gives a nice wide barrier, but for some reason the barrier is disintegrating on our slides so I get holes in the lines I draw. I have had a similar problem before, usually because the slide wasn't dry enough when I was drawing the barrier or I was immersing the slide before the barrier had a chance to solidify. However, the problem has never been this bad. I am wondering if my current batch of pap pens is defective. Patrick CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information protected by law. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Anyone using Section Transfer System by Microm?
Tim, HM340 + STS now successfully uses in many Russians hisology labs and techs loves them. STS have the one disadvantage: is need to much time for cleaning and maintenance vs waterbath. Also it is necessary some skills for using it. Sincerely, Maxim Peshkov, Russia, Taganrog. mailto:maxim...@mail.ru ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Has anyone had any problems with the Vector Immuedge pap pen?
Hi Patrick, yes, I had a similar problem with the Immedge Pap pen. I took very long before I could immerse the sections again with buffer and the barrier still collapsed later on. I used a quite old one that I found in the lab so I guess they stop functioning after a while. Klaus Klaus Kruttwig Department of Cell and Tissue Biology University of California, San Francisco HSW601 From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] on behalf of Lewis, Patrick [patrick.le...@seattlechildrens.org] Sent: Thursday, November 14, 2013 10:19 AM To: 'Histonet@lists.utsouthwestern.edu' Subject: [Histonet] Has anyone had any problems with the Vector Immuedge pap pen? Hi everyone, Has anyone had any problems with the Immedge Pap pen from Vector. Cat # H-4000 It gives a nice wide barrier, but for some reason the barrier is disintegrating on our slides so I get holes in the lines I draw. I have had a similar problem before, usually because the slide wasn't dry enough when I was drawing the barrier or I was immersing the slide before the barrier had a chance to solidify. However, the problem has never been this bad. I am wondering if my current batch of pap pens is defective. Patrick CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information protected by law. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: ImmEdge Pap Pen problems
The problem is using the pen without redistributing the liquid and solid contents in the pen. Years ago, the Vector technical service rep said to shake the daylights out of the pen. We use a vortex mixer, at highest speed and press hard, to make sure the contents were well mixed before use. We then have no problems of the goo lifting off the slide when in buffer or using a solvent fixative for frozen sections after applying the barrier. We had pens that worked for years as long as we shook (or vortexed) them well before applying the barrier around sections. We used the pens for both deparaffinized slides (well dried around section) and air dried frozen section (on dry slides). Good luck Gayle Callis HTL/HT/MT(ASCP) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
