[Histonet] help ! Dako Seymour labeling software - autostainer plus
Hello dear histoneters First let me thank you all for your help for lost password (autostainer plus software) - a - a tip is ok. I am begining to test the autostainer and now eveything seems fine (there was a stuck tube but I could settle the problem) _Corrupted files : Dako Seymour software (TLP 2742 printer)_ Autostainer plus can print labels directly from main software. But printing labels can be done from another computer if you use a another provided software (Dako Seymour labeling software) My software dako Seymour 5.0 is on two floppies and two compressed files are corrupted */VB40016.DL_/* (on first floppy) and */ARW01DN0.IC_/* (on second floppy) _It would be great if someone could send me by mail the two missing working files_ (compressed format, exactly as I have written them here would be better, but may be uncompressed files that are in the computer could work - these files are not very heavy and can easily be sent) I asked Dako France but they dont have anymore the floppies of the Dako Seymour labeling software (5.0) _Does someone have a list or a link of best protocols (procedures) for the machine ?_ have a nice day Best regards R Wild ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Productivity
Our unit of measure is billable test. But, my director adds in extra (not sure how) for manual vs. automation in the clinical lab. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Heckford, Karen - SMMC-SF Sent: Monday, August 20, 2012 1:43 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Productivity Good Morning, I know we have been over this before, how does your lab measure productivity? Are you going by block and slide or by CPT code? I have been told by the Suits that we can only measure productivity by per specimen or CPT code with no weight and value. As you know when you get a big case you can have a bunch of blocks on it therefore a bunch of slides. I do everything from GI specimens to big breast cases. They will tell me that our productivity is down and we need to cut hours. I just do not know exactly how to drill it in to them that things are not always as they appear on paper. I am talking in circles until I am blue in the face...AUGH!! Karen Heckford HT ASCP CE Lead Histology Technician St. Mary's Medical Center 450 Stanyan St. San Francisco, Ca. 94117 415-668-1000 ext. 6167 karen.heckf...@dignityhealth.org Caution: This email message, including all content and attachments, is CONFIDENTIAL and may be of a nature that is LEGALLY PRIVILEGED. The information contained in this email message is intended only for the use of the recipient(s) named above. If the reader of this message is not the intended recipient or an agent responsible for delivering it to the intended recipient, you have received this document in error. Any further review, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please notify us immediately by reply email. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Disclaimer: This electronic message may contain information that is Proprietary, Confidential, or legally privileged or protected. It is intended only for the use of the individual(s) and entity named in the message. If you are not an intended recipient of this message, please notify the sender immediately and delete the material from your computer. Do not deliver, distribute or copy this message and do not disclose its contents or take any action in reliance on the information it contains. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Formalin and Operating Rooms
Debra and others: All of our specimens are sent fresh because surgery says formalin cannot be in the operating rooms. I'm told it comes from AORN, which is the association for operating room nurses. Hazel Horn Supervisor of Histology/Autopsy/Transcription Anatomic Pathology Arkansas Children's Hospital 1 Children's Way | Slot 820| Little Rock, AR 72202 501.364.4240 direct | 501.364.1302 office | 501.364.1241 fax hor...@archildrens.org archildrens.org 100 YEARS YOUNG! JOIN THE PARTY AT ach100.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Debra Siena Sent: Monday, August 20, 2012 5:25 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Formalin and Operating Rooms Hi All, I would like to ask if anyone has heard of any new regulations or laws that state that the Operating Room can't have formalin available in the room so that they can place formalin onto the sample right away? I was wondering if anyone has heard of this, if you could tell me more about where it is coming from so that I can access a copy of it. We have had some inquiries and I have not heard of this. I appreciate any help that you can give and sorry, that I don't have more information. Best wishes. Debbie Siena, HT(ASCP)QIHC StatLab Medical Products Technical Support Manager 407 Interchange Street | McKinney, TX 75071 t: 800.442.3573 ext. 229 | f: 972.436.1369 dsi...@statlab.com | www.statlab.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ** The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Renal Cell Carcinoma Poll
All, What is your favorite Renal Cell Carcinoma Antibody? I'm using the Leica Bond stainer with Leica's detection am looking for the best fit. In other words, the antibody that I am using is working, but I want it to look better. Any responses would be greatly appreciated. Glen Dawson BS, HT(ASCP), QIHC IHC Technical Specialist Janesville, WI ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Formalin and Operating Rooms
I know of no rules, though there may well be. I do know that several years ago a patient was injected (IV)with formalin in the OR. Results were NOT good. I know that the OR in the place I was working at the time would no longer allow for formalin in the OR suite. The tissues were put in formalin in a separate area - by OR staff. Bill -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Paula Sicurello Sent: Monday, August 20, 2012 6:05 PM To: Debra Siena Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Formalin and Operating Rooms I would be interested in the replies as well. To add to that, I've heard the same thing about glutartaldehyde, do the same rules apply? Thanks, Paula -- Paula Sicurello, HTL (ASCP) Supervisor, Clinical Electron Microscopy Laboratory Duke University Health System Rm.#251M, Duke South, Green Zone Durham, North Carolina 27710 P: 919.684.2091 On Mon, Aug 20, 2012 at 6:25 PM, Debra Siena dsi...@statlab.com wrote: Hi All, I would like to ask if anyone has heard of any new regulations or laws that state that the Operating Room can't have formalin available in the room so that they can place formalin onto the sample right away? I was wondering if anyone has heard of this, if you could tell me more about where it is coming from so that I can access a copy of it. We have had some inquiries and I have not heard of this. I appreciate any help that you can give and sorry, that I don't have more information. Best wishes. Debbie Siena, HT(ASCP)QIHC StatLab Medical Products Technical Support Manager 407 Interchange Street | McKinney, TX 75071 t: 800.442.3573 ext. 229 | f: 972.436.1369 dsi...@statlab.com | www.statlab.com ___ 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 This electronic mail and any attached documents are intended solely for the named addressee(s) and contain confidential information. If you are not an addressee, or responsible for delivering this email to an addressee, you have received this email in error and are notified that reading, copying, or disclosing this email is prohibited. If you received this email in error, immediately reply to the sender and delete the message completely from your computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Formalin and Operating Rooms
I also have not heard of any recent rule changes. Being aware of the formalin mishap in OR, our hospital has been using for over 20 years, a light blue tinged 10%NBF that the supplier makes up special. The blue is actually just a few drops of food coloring that the manufacturer adds, and the blue does NOT carry over through processing and staining. I am also aware that we are not the only hospital using this blue formalin. There is no confusion for the OR personnel when they grab the appropriate container (NBF v/s saline). hope this helps, Lynette From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] on behalf of O'Donnell, Bill [billodonn...@catholichealth.net] Sent: Tuesday, August 21, 2012 9:05 AM To: Paula Sicurello; Debra Siena Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Formalin and Operating Rooms I know of no rules, though there may well be. I do know that several years ago a patient was injected (IV)with formalin in the OR. Results were NOT good. I know that the OR in the place I was working at the time would no longer allow for formalin in the OR suite. The tissues were put in formalin in a separate area - by OR staff. Bill -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Paula Sicurello Sent: Monday, August 20, 2012 6:05 PM To: Debra Siena Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Formalin and Operating Rooms I would be interested in the replies as well. To add to that, I've heard the same thing about glutartaldehyde, do the same rules apply? Thanks, Paula -- Paula Sicurello, HTL (ASCP) Supervisor, Clinical Electron Microscopy Laboratory Duke University Health System Rm.#251M, Duke South, Green Zone Durham, North Carolina 27710 P: 919.684.2091 On Mon, Aug 20, 2012 at 6:25 PM, Debra Siena dsi...@statlab.com wrote: Hi All, I would like to ask if anyone has heard of any new regulations or laws that state that the Operating Room can't have formalin available in the room so that they can place formalin onto the sample right away? I was wondering if anyone has heard of this, if you could tell me more about where it is coming from so that I can access a copy of it. We have had some inquiries and I have not heard of this. I appreciate any help that you can give and sorry, that I don't have more information. Best wishes. Debbie Siena, HT(ASCP)QIHC StatLab Medical Products Technical Support Manager 407 Interchange Street | McKinney, TX 75071 t: 800.442.3573 ext. 229 | f: 972.436.1369 dsi...@statlab.com | www.statlab.com ___ 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 This electronic mail and any attached documents are intended solely for the named addressee(s) and contain confidential information. If you are not an addressee, or responsible for delivering this email to an addressee, you have received this email in error and are notified that reading, copying, or disclosing this email is prohibited. If you received this email in error, immediately reply to the sender and delete the message completely from your computer 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
RE: [Histonet] DAB precipitate on H. pylori
We never had a problem until they changed AB's on us. We sent them slides procedures etc to test but in the mean time we tried Cell Marques AB and it was clean. Problem solved. Now we all need to demand that Ventana switch back to Cell Marques H pylori so that we do not have to pay for those #@*% prep kits. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Vanessa Perez Sent: Monday, August 20, 2012 2:11 PM To: Dorothy Glass; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] DAB precipitate on H. pylori Are you using the ventana h. pylori? If you are we are having that problem with some of our satellite labs. Ventana said its an issue with that new antibody and some labs but not all are experiencing this. Have you called it in the Customer support??? Vanessa Perez Garcia Histology Supervisor Pathology Reference Lab 210-892-3746 210-892-3732 vpe...@pathreflab.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Dorothy Glass Sent: Monday, August 20, 2012 1:13 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] DAB precipitate on H. pylori I have been getting something on some slides, not all that looks like bacteria but it is not. It looks like Dab precipitate. It looks like the Dab is breaking down, but only noticeable on the stains for H. pylori. I wash my water bath, and changes my blades. It not occurs all the time, but it has happened about four times. Could it be the water not being filtered properly. We have a water filtering system, but it has not been working well lately. Would that have anything to do with the brown patches on H. pylori? ___ 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] Position - Snr Histotechnician
We have a vacancy for a Senior Histotechnican. The Senior Histotechnician is responsible for performing a wide variety of histotechnological testing including, but not limited to, microtomy, cryotomy, complex special stains, IF, IHC, enzyme histochemistry and electron microscopy. The potential exists to become involved in research projects within the department. A working knowledge of Lean principles is useful. Theoretical knowledge as well as high quality practical ability is required. Candidates with expertise in Immunohistochemistry and/or Electron Microscopy preferred. Experience in pediatrics desirable. Registered HT(ASCP) or equivalent, with a minimum of 5 years histopathology experience in a similar laboratory Please apply on our web-site, and/or contact Sue Doud (614-355-41-5) or myself for more details Thanks Ronnie Houston, MS HT(ASCP)QIHC Anatomic Pathology Manager ChildLab, a Division of Nationwide Children's Hospital www.childlab.com 700 Children's Drive Columbus, OH 43205 (P) 614-722-5450 (F) 614-722-2899 ronald.hous...@nationwidechildrens.orgmailto:ronald.hous...@nationwidechildrens.org www.NationwideChildrens.orghttp://www.nationwidechildrens.org/ One person with passion is better than forty people merely interested. ~ E.M. Forster - Confidentiality Notice: The following mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. The recipient is responsible to maintain the confidentiality of this information and to use the information only for authorized purposes. If you are not the intended recipient (or authorized to receive information for the intended recipient), you are hereby notified that any review, use, disclosure, distribution, copying, printing, or action taken in reliance on the contents of this e-mail is strictly prohibited. If you have received this communication in error, please notify us immediately by reply e-mail and destroy all copies of the original message. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Formalin and Operating Rooms
Here is a notorious case from my days working in a Ft. Lauderdale suburban hospital. Along with Bill's experiences we took formalin out of the OR after this was reported, but it wasn't a requirement as far as I remember. http://www.nytimes.com/1985/03/10/us/one-death-many-questions-in-miami.html?pagewanted=all Brett M. Connolly, Ph.D. Principal Scientist, Imaging Dept. Merck Co., Inc. PO Box 4, WP-44K West Point, PA 19486 brett_conno...@merck.com T- 215-652-2501 F- 215-993-6803 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of O'Donnell, Bill Sent: Tuesday, August 21, 2012 9:05 AM To: Paula Sicurello; Debra Siena Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Formalin and Operating Rooms I know of no rules, though there may well be. I do know that several years ago a patient was injected (IV)with formalin in the OR. Results were NOT good. I know that the OR in the place I was working at the time would no longer allow for formalin in the OR suite. The tissues were put in formalin in a separate area - by OR staff. Bill -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Paula Sicurello Sent: Monday, August 20, 2012 6:05 PM To: Debra Siena Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Formalin and Operating Rooms I would be interested in the replies as well. To add to that, I've heard the same thing about glutartaldehyde, do the same rules apply? Thanks, Paula -- Paula Sicurello, HTL (ASCP) Supervisor, Clinical Electron Microscopy Laboratory Duke University Health System Rm.#251M, Duke South, Green Zone Durham, North Carolina 27710 P: 919.684.2091 On Mon, Aug 20, 2012 at 6:25 PM, Debra Siena dsi...@statlab.com wrote: Hi All, I would like to ask if anyone has heard of any new regulations or laws that state that the Operating Room can't have formalin available in the room so that they can place formalin onto the sample right away? I was wondering if anyone has heard of this, if you could tell me more about where it is coming from so that I can access a copy of it. We have had some inquiries and I have not heard of this. I appreciate any help that you can give and sorry, that I don't have more information. Best wishes. Debbie Siena, HT(ASCP)QIHC StatLab Medical Products Technical Support Manager 407 Interchange Street | McKinney, TX 75071 t: 800.442.3573 ext. 229 | f: 972.436.1369 dsi...@statlab.com | www.statlab.com ___ 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 This electronic mail and any attached documents are intended solely for the named addressee(s) and contain confidential information. If you are not an addressee, or responsible for delivering this email to an addressee, you have received this email in error and are notified that reading, copying, or disclosing this email is prohibited. If you received this email in error, immediately reply to the sender and delete the message completely from your computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Notice: This e-mail message, together with any attachments, contains information of Merck Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates Direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] annoying crystals on sections
we are using a Sakura DRS2000 and we are 3 x 3 minutes in xylene and we have been keeping it fresh. The staining is good now but we still see the crystals. If it were paraffin we should see unstained spots on the slide I think. I have gone to an aqueous 1% HCl today after hematoxylin for regression and that seems to be cleaning them up on most of the slides. I cleaned out some of the plumbing and cleaned some calcium out of the pipes. We are using Harris hematoxylin that we purchase. We have a tried different counterstains but it seems to make no difference. We are using a Sakura tissue processor for overnight processing of cassettes. the embedding is going good and we get nice flat thin sections. We are fixing tissues with neutral phosphate buffered formalin but still see some formalin pigment. We are cleaning that up with picric acid in etoh. We find we still need that and the formalin pigment is brown to dark brown. These problem crystals are round irregular to rhomboidal some times sort of large and flat about the size of a cell and they are clear. I thought they were formalin pigment at first and fiddled with the Picric Acid, and tried Ammonia in Alcohol to get rid of formalin pigment and finally decided that it was not formalin pigment. I thought it might be something from Scott's tap water (Mg++) so i dropped that and tried bluing with NH4+ and it didnt help any. I tried blueing just with tap water. Nice result but still the crystals. The aqueous HCl seems to be working and is not harming the nuclei so I may have a sort of solution and am calling it calcium crystals in the water until I know better. I may look for some sort of filter to put in the water line. E Wayne Johnson DVM Enruikang Ag Tech MOA Feed Industry Centre China Agriculture University Beijing On 8/21/2012 7:51 PM, Debra Siena wrote: could it be paraffin? Debbie Siena HT(ASCP)QIHC Technical Manager | StatLab Medical Products 407 Interchange St. | McKinney, TX 75071 Direct: 972-436-1010 x229 | Fax: 972-436-1369 dsi...@statlab.com | www.statlab.com - Original Message - From: e...@pigsqq.org [mailto:e...@pigsqq.org] Sent: Tuesday, August 21, 2012 05:46 AM To: Debra Siena Subject: Re: [Histonet] annoying crystals on sections We just now ran the statlab version protocol from StatLab's website, using an alcoholic eosin. No doubt that gives a stronger brighter red stain. However we still see those crystals! We are suspecting a water problem. I have been dismantling some of the plumbing and getting some Ca crystals out of the pipes. We are manually coverslipping. ---Original Message--- From: Debra Sienadsi...@statlab.com To: 'e...@pigsqq.org'e...@pigsqq.org Subject: Re: [Histonet] annoying crystals on sections Sent: Aug 21 '12 07:46 Is your eosin alcoholic or aqueous? What is your staining protocol and what reagents are you using? This information would be most helpful. Also are the sections paraffin embedded and routinely processed in tissue processor? Debbie Siena HT(ASCP)QIHC Technical Manager | StatLab Medical Products 407 Interchange St. | McKinney, TX 75071 Direct: 972-436-1010 x229 | Fax: 972-436-1369 dsi...@statlab.com | www.statlab.com - Original Message - From: E. Wayne Johnson [mailto:e...@pigsqq.org] Sent: Monday, August 20, 2012 06:20 PM To: histonet@lists.utsouthwestern.eduhistonet@lists.utsouthwestern.edu Subject: [Histonet] annoying crystals on sections We are having problems with crystals precipitated on our slides which are HE stains on tissues from pigs. Tissues are fixed in buffered formalin. We had trouble months ago with formalin pigment and we had resolved that by using ammonia in EtOH or picric acid in EtOH. Sometimes we receive fixed samples from the field that are not buffered but presently all of our tissues are fixed in neutral phosphate buffered formalin. We moved the Sakura autostainer to a different location under a fume hood on a different floor of the building to get the solvent odor out of our work area. Immediately we began to see a tremendous degradation in slide quality due to what we initially thought was formalin pigment. We have changed all of the solutions and all of the stains. We find that if we use Milli-q water instead of tap water for rinsing (done by hand in that case) we dont see the crystals, but the eosin staining quality is not acceptable after rinsing in the acidic (ph ~5) Milli-Q water. Our tap water is neutral to slightly alkaline and is very hard with calcium. We do all sorts of tissues for diagnosis of pig diseases. Sometimes the slides are quite acceptable but sometimes particularly when looking at small intestine, the crystals are very annoying. The crystals occur randomly on the slide except that there is a tendency for them to be centered on nuclei particularly in intestinal epithelium. The crystals
RE: [Histonet] annoying crystals on sections
Hi Wayne, I had lots of problems with round irregular crystals, but have greatly improved my slides by limiting baking at 57 degrees to 1/2 hour. We use a paraffin with plastic in the mix, and I think the plastic globs up under some conditions, like no or not enough xylene to dissolve it out. Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of E. Wayne Johnson Sent: Tuesday, August 21, 2012 8:09 AM To: Debra Siena; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] annoying crystals on sections we are using a Sakura DRS2000 and we are 3 x 3 minutes in xylene and we have been keeping it fresh. The staining is good now but we still see the crystals. If it were paraffin we should see unstained spots on the slide I think. I have gone to an aqueous 1% HCl today after hematoxylin for regression and that seems to be cleaning them up on most of the slides. I cleaned out some of the plumbing and cleaned some calcium out of the pipes. We are using Harris hematoxylin that we purchase. We have a tried different counterstains but it seems to make no difference. We are using a Sakura tissue processor for overnight processing of cassettes. the embedding is going good and we get nice flat thin sections. We are fixing tissues with neutral phosphate buffered formalin but still see some formalin pigment. We are cleaning that up with picric acid in etoh. We find we still need that and the formalin pigment is brown to dark brown. These problem crystals are round irregular to rhomboidal some times sort of large and flat about the size of a cell and they are clear. I thought they were formalin pigment at first and fiddled with the Picric Acid, and tried Ammonia in Alcohol to get rid of formalin pigment and finally decided that it was not formalin pigment. I thought it might be something from Scott's tap water (Mg++) so i dropped that and tried bluing with NH4+ and it didnt help any. I tried blueing just with tap water. Nice result but still the crystals. The aqueous HCl seems to be working and is not harming the nuclei so I may have a sort of solution and am calling it calcium crystals in the water until I know better. I may look for some sort of filter to put in the water line. E Wayne Johnson DVM Enruikang Ag Tech MOA Feed Industry Centre China Agriculture University Beijing On 8/21/2012 7:51 PM, Debra Siena wrote: could it be paraffin? Debbie Siena HT(ASCP)QIHC Technical Manager | StatLab Medical Products 407 Interchange St. | McKinney, TX 75071 Direct: 972-436-1010 x229 | Fax: 972-436-1369 dsi...@statlab.com | www.statlab.com - Original Message - From: e...@pigsqq.org [mailto:e...@pigsqq.org] Sent: Tuesday, August 21, 2012 05:46 AM To: Debra Siena Subject: Re: [Histonet] annoying crystals on sections We just now ran the statlab version protocol from StatLab's website, using an alcoholic eosin. No doubt that gives a stronger brighter red stain. However we still see those crystals! We are suspecting a water problem. I have been dismantling some of the plumbing and getting some Ca crystals out of the pipes. We are manually coverslipping. ---Original Message--- From: Debra Sienadsi...@statlab.com To: 'e...@pigsqq.org'e...@pigsqq.org Subject: Re: [Histonet] annoying crystals on sections Sent: Aug 21 '12 07:46 Is your eosin alcoholic or aqueous? What is your staining protocol and what reagents are you using? This information would be most helpful. Also are the sections paraffin embedded and routinely processed in tissue processor? Debbie Siena HT(ASCP)QIHC Technical Manager | StatLab Medical Products 407 Interchange St. | McKinney, TX 75071 Direct: 972-436-1010 x229 | Fax: 972-436-1369 dsi...@statlab.com | www.statlab.com - Original Message - From: E. Wayne Johnson [mailto:e...@pigsqq.org] Sent: Monday, August 20, 2012 06:20 PM To: histonet@lists.utsouthwestern.eduhistonet@lists.utsouthwestern.edu Subject: [Histonet] annoying crystals on sections We are having problems with crystals precipitated on our slides which are HE stains on tissues from pigs. Tissues are fixed in buffered formalin. We had trouble months ago with formalin pigment and we had resolved that by using ammonia in EtOH or picric acid in EtOH. Sometimes we receive fixed samples from the field that are not buffered but presently all of our tissues are fixed in neutral phosphate buffered formalin. We moved the Sakura autostainer to a different location under a fume hood on a different floor of the building to get the solvent odor out of our work area. Immediately we began to see a tremendous degradation in slide quality due to what we initially thought was formalin pigment. We have changed all of the solutions and all of the stains. We find that if we
Re: [Histonet] annoying crystals on sections
Obviously, it's your water, if you don't see the crystals after changing your water. If the acidic pH of the Milli-q water is leaching out your eosin, then adjust the pH of the water to 7.0 before using it to rinse your slides. Sincerely, Jay A. Lundgren, M.S., HTL (ASCP) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Formalin and Operating Rooms
Johnson Johnson just a few days ago announced that they're going to take formaldehyde out of baby shampoo! No wonder I can't get my baby's hair squeeky clean anymore! How much formalin do I need to add to replace the formalin they took out? Sincerely? Jay A. Lundgren, M.S., HTL (ASCP) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Accessions/ lean
My facility currently takes specimens out of plastic bags, accessions the case, and then stuffs the specimens back into the plastic bags. One of the residents mentioned that other places put the specimens into plastic bins for the grossers. I have been asked to evaluate the feasability of this, but I have never seen it done, don't know how the bins would stack, how many different sizes would be needed, how much room it would take up, etc. If anyone that does this can comment or better yet could provide a photo(s) of the bin set-up at accessioning/grossing that would be great. Thanks in advance!! Nisha Lead histology technian Robert Wood Johnson University Hospital Sent from my iPhone ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Training -LIS
Hello HistonettersI am wondering if anyone can direct me to some good training courses or sources for IT training, but specifically in areas that would be of good use for LIS. I have taken some health informatics courses with SQL, some LOINC,and EMR topics and I am looking to get into these studies a little further. I am not really looking to go into a formal degree program at this time, I am wanting to learn more but only as a part-time study venture. Thanks for your input.Joelle Joelle Weaver MAOM, HTL (ASCP) QIHC ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Open jobs - including Supervisor Manager
Hello my fellow 'netters! We have another bunch of openings-- Routine bench-- Day, afternoon and night shift -- all over the country and registered and non-registered openings are available. Supervisor- Several openings includind a night shift position Manager- One big job and several solo positions. Call for more information-- be ready with your most recent resume and some will require education transcripts even if you've not finished the degree If what we have doesn't suit, we'll go looking for you! The conversation is with me, a working histotech--so it is quick and usually kinda fun. Thank you! In service! Cheryl Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one GREAT Tech at a time. 281.852.9457 Office 800.756.3309 Phone Fax ad...@fullstaff.org Sign up for the FREE newsletter AP News--updates, tricks of the trade and current issues for Anatomic Pathology Clinical Labs. Send a 'subscribe' request to apn...@fullstaff.org. Please include your name and specialty in the body of the email. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Accessions/ lean
Regardless of what you end doing, NEVER touch anything twice to return to the same container it came. The idea is that every time you touch a specimen you add value to it. This means that the specimen should fall into a chain of work, into a forward flow that makes the specimen one step closer to finish every time you handle it. Taken a specimen from a plastic bag to put it AGAIN into the same plastic bag is anathema from the Lean point of view. Study your flow and try that your specimen keeps marching forward with every step you subject it to. René J. From: Nisha monishapah...@gmail.com To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Sent: Tuesday, August 21, 2012 1:32 PM Subject: [Histonet] Accessions/ lean My facility currently takes specimens out of plastic bags, accessions the case, and then stuffs the specimens back into the plastic bags. One of the residents mentioned that other places put the specimens into plastic bins for the grossers. I have been asked to evaluate the feasability of this, but I have never seen it done, don't know how the bins would stack, how many different sizes would be needed, how much room it would take up, etc. If anyone that does this can comment or better yet could provide a photo(s) of the bin set-up at accessioning/grossing that would be great. Thanks in advance!! Nisha Lead histology technian Robert Wood Johnson University Hospital Sent from my iPhone ___ 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] Gloves
Hi, what type of gloves are you wearing when handling xylene? Do you double glove? Cordially, Pam ~ Pam Richardson Clinical Manager Gundersen Lutheran Laboratory Email: pkric...@gundluth.org Phone: 608 775-4133 Fax: 608 775-6136 Interdepartmental Mail Stop: H04-007 E-visit us at: http://www.gundluth.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Gloves
The best type is the one used to handle paint removers (paint thinners). René J. From: Richardson, Pam K pkric...@gundluth.org To: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu Sent: Tuesday, August 21, 2012 3:52 PM Subject: [Histonet] Gloves Hi, what type of gloves are you wearing when handling xylene? Do you double glove? Cordially, Pam ~ Pam Richardson Clinical Manager Gundersen Lutheran Laboratory Email: pkric...@gundluth.org Phone: 608 775-4133 Fax: 608 775-6136 Interdepartmental Mail Stop: H04-007 E-visit us at: http://www.gundluth.org ___ 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] Accessions/ lean
We use baskets here. They get them from office depot. Put each case in one basket and then put on a cart to deliver to the grosser. For big specimens that don't fit we just set the container on the grossing table. Vanessa Perez Garcia Histology Supervisor Pathology Reference Lab 210-892-3746 210-892-3732 vpe...@pathreflab.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Nisha Sent: Tuesday, August 21, 2012 12:32 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Accessions/ lean My facility currently takes specimens out of plastic bags, accessions the case, and then stuffs the specimens back into the plastic bags. One of the residents mentioned that other places put the specimens into plastic bins for the grossers. I have been asked to evaluate the feasability of this, but I have never seen it done, don't know how the bins would stack, how many different sizes would be needed, how much room it would take up, etc. If anyone that does this can comment or better yet could provide a photo(s) of the bin set-up at accessioning/grossing that would be great. Thanks in advance!! Nisha Lead histology technian Robert Wood Johnson University Hospital Sent from my iPhone ___ 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: Gloves and Xylene
For ordinary applications like manual staining, nitrile gloves work fine. I don't double glove, just change gloves ad lib. Jerry From: pkric...@gundluth.org To: histonet@lists.utsouthwestern.edu Date: Tue, 21 Aug 2012 19:52:50 + Subject: [Histonet] Gloves Hi, what type of gloves are you wearing when handling xylene? Do you double glove? Cordially, Pam ~ Pam Richardson Clinical Manager Gundersen Lutheran Laboratory Email: pkric...@gundluth.org Phone: 608 775-4133 Fax: 608 775-6136 Interdepartmental Mail Stop: H04-007 E-visit us at: http://www.gundluth.org ___ 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] Gloves
Hi Pam: I use nitrile gloves that we get from our Central Supply Room when handling xylene and cover-slipping by hand. They are Kimberly-Clark Sterling Nitrile Powder-Free Exam Gloves. I use just a single glove rather than double-gloving, and that seems to do the trick. The glove usually does not break. However, now that you mention it, I wonder if a single layer really totally prevents penetration of xylene or not. Tim Tim Wheelock Harvard Brain Bank McLean Hospital Belmont, MA 02478 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] cryosections with DiD labeling
We are beginning a retrograde tracing study in rats. We injected DiD into the rats and in 4 weeks we will perfuse and harvest the brainstems. My question is, is vibratome sectioning better than cryostat sectioning? The literature that accompanied the dye said either is an option, but some have noted dye leaching out when cyrostat sectioned. How common is this problem? If vibratome is the better choice, is there a good embedding protocol? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Gloves
We use nitrile when handling everything except acetone - then we use latex. Tresa -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Richardson, Pam K Sent: Tuesday, August 21, 2012 1:53 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Gloves Hi, what type of gloves are you wearing when handling xylene? Do you double glove? Cordially, Pam ~ Pam Richardson Clinical Manager Gundersen Lutheran Laboratory Email: pkric...@gundluth.org Phone: 608 775-4133 Fax: 608 775-6136 Interdepartmental Mail Stop: H04-007 E-visit us at: http://www.gundluth.org ___ 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: Gloves
Pam I hand coverslip and have found in time most of the nitrile gloves break down and get real soft. However, by accident our purchasing department ordered SemperSure Nitrile gloves (tested for use with chemothepary drugs) and these are the only gloves that I found that don't get soft and break down. There is 200 gloves in the box and are true to size They are by Sempermed the order number for the small is SUNF202 Kate Kate Mendell Histopathology/Lab Manager HOWARD S. GOLDBERG, M.D., INC 990 Paradise Road Swampscott, MA 01907 TEL: 781.595.0151 FAX: 781.592.6780 kmend...@goldbergmd.net www.cosmesticdermcenter.com PRIVACY NOTICE: This e-mail message may contain confidential patient or other information belonging to the sender that is legally privileged. This information is intended only for the use of the individual or authorized entity named above. The authorized recipient of this patient or other confidential information is prohibited from disclosing the information to any other party. If you have received this message in error, please notify the sender immediately and delete. Please keep any information you may have viewed confidential. From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Richardson, Pam K [pkric...@gundluth.org] Sent: Tuesday, August 21, 2012 3:52 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Gloves Hi, what type of gloves are you wearing when handling xylene? Do you double glove? Cordially, Pam ~ Pam Richardson Clinical Manager Gundersen Lutheran Laboratory Email: pkric...@gundluth.org Phone: 608 775-4133 Fax: 608 775-6136 Interdepartmental Mail Stop: H04-007 E-visit us at: http://www.gundluth.org ___ 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: annoying crystals on sections
Hi Wayne Histonet My guess is that Wayne's crystals are Calcium phosphate - the calcium from the hard tap water, as described, and phosphate from phosphate buffer (sodium or potassium) in the solution immediately preceding the tapwater rinse. The variation in crystal deposition would then be in the degree each tissue/ organelle tends to carry over the phosphate into the tapwater wash. Just mix drops of the solutions together on a slide and see if crystals form. A brief deionized rinse followed by tapwater should first remove the phosphate ( crystals) and then allow the desired blueing. Alternatively, substitute TRIS or similar as the buffer in the preceding step and go directly to tapwater. If you have valuable sections with crystals on them, you should be able to chelate away the deposits in an EDTA solution, then restain as needed. all the best! -David == David A. Wright, Ph.D. University of Chicago Section of Neurosurgery Original message Date: Tue, 21 Aug 2012 09:44:26 -0500 (CDT) Subject: Histonet Digest, Vol 105, Issue 25 Message: 12 Date: Tue, 21 Aug 2012 07:20:06 +0800 From: E. Wayne Johnson e...@pigsqq.org Subject: [Histonet] annoying crystals on sections We are having problems with crystals precipitated on our slides which are HE stains on tissues from pigs. Tissues are fixed in buffered formalin. We had trouble months ago with formalin pigment and we had resolved that by using ammonia in EtOH or picric acid in EtOH. Sometimes we receive fixed samples from the field that are not buffered but presently all of our tissues are fixed in neutral phosphate buffered formalin. We moved the Sakura autostainer to a different location under a fume hood on a different floor of the building to get the solvent odor out of our work area. Immediately we began to see a tremendous degradation in slide quality due to what we initially thought was formalin pigment. We have changed all of the solutions and all of the stains. We find that if we use Milli-q water instead of tap water for rinsing (done by hand in that case) we don't see the crystals, but the eosin staining quality is not acceptable after rinsing in the acidic (ph ~5) Milli-Q water. Our tap water is neutral to slightly alkaline and is very hard with calcium. We do all sorts of tissues for diagnosis of pig diseases. Sometimes the slides are quite acceptable but sometimes particularly when looking at small intestine, the crystals are very annoying. The crystals occur randomly on the slide except that there is a tendency for them to be centered on nuclei particularly in intestinal epithelium. The crystals are birefringent in polarized light but seem to be generally clear not dark like the formalin pigment we had seen before. Neither ammonia nor picric acid remove these, and now if we use alcoholic ammonia to treat the slides, the slides come out too blue. Our slides are cut at 4 to 5 microns. Brain has the least problem, small intestine seems worst. We have gone back and cut some blocks that previously stained beautifully with no pigment or precipitate problems and those slides also now have the same problem, either crystals if washed with tap water, or poor eosin staining if rinsed with MilliQ water. Our next step is to examine the slides microscopically at every step and try to find at which step the problem is occurring. Any thoughts or similar experiences? E. Wayne Johnson, DVM Enruikang AgTech MOA Feed Industry Centre Beijing -- ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: annoying crystals on sections
Thanks all for many useful and helpful suggestions and interesting anecdotes. E. Wayne Johnson, DVM Enruikang AgTech MOA Feed Industry Centre China Agricultural University Beijing On 8/22/2012 5:27 AM, David A. Wright wrote: Hi Wayne Histonet My guess is that Wayne's crystals are Calcium phosphate - the calcium from the hard tap water, as described, and phosphate from phosphate buffer (sodium or potassium) in the solution immediately preceding the tapwater rinse. The variation in crystal deposition would then be in the degree each tissue/ organelle tends to carry over the phosphate into the tapwater wash. Just mix drops of the solutions together on a slide and see if crystals form. A brief deionized rinse followed by tapwater should first remove the phosphate ( crystals) and then allow the desired blueing. Alternatively, substitute TRIS or similar as the buffer in the preceding step and go directly to tapwater. If you have valuable sections with crystals on them, you should be able to chelate away the deposits in an EDTA solution, then restain as needed. all the best! -David == David A. Wright, Ph.D. University of Chicago Section of Neurosurgery Original message Date: Tue, 21 Aug 2012 09:44:26 -0500 (CDT) Subject: Histonet Digest, Vol 105, Issue 25 Message: 12 Date: Tue, 21 Aug 2012 07:20:06 +0800 From: E. Wayne Johnsone...@pigsqq.org Subject: [Histonet] annoying crystals on sections We are having problems with crystals precipitated on our slides which are HE stains on tissues from pigs. Tissues are fixed in buffered formalin. We had trouble months ago with formalin pigment and we had resolved that by using ammonia in EtOH or picric acid in EtOH. Sometimes we receive fixed samples from the field that are not buffered but presently all of our tissues are fixed in neutral phosphate buffered formalin. We moved the Sakura autostainer to a different location under a fume hood on a different floor of the building to get the solvent odor out of our work area. Immediately we began to see a tremendous degradation in slide quality due to what we initially thought was formalin pigment. We have changed all of the solutions and all of the stains. We find that if we use Milli-q water instead of tap water for rinsing (done by hand in that case) we don't see the crystals, but the eosin staining quality is not acceptable after rinsing in the acidic (ph ~5) Milli-Q water. Our tap water is neutral to slightly alkaline and is very hard with calcium. We do all sorts of tissues for diagnosis of pig diseases. Sometimes the slides are quite acceptable but sometimes particularly when looking at small intestine, the crystals are very annoying. The crystals occur randomly on the slide except that there is a tendency for them to be centered on nuclei particularly in intestinal epithelium. The crystals are birefringent in polarized light but seem to be generally clear not dark like the formalin pigment we had seen before. Neither ammonia nor picric acid remove these, and now if we use alcoholic ammonia to treat the slides, the slides come out too blue. Our slides are cut at 4 to 5 microns. Brain has the least problem, small intestine seems worst. We have gone back and cut some blocks that previously stained beautifully with no pigment or precipitate problems and those slides also now have the same problem, either crystals if washed with tap water, or poor eosin staining if rinsed with MilliQ water. Our next step is to examine the slides microscopically at every step and try to find at which step the problem is occurring. Any thoughts or similar experiences? E. Wayne Johnson, DVM Enruikang AgTech MOA Feed Industry Centre Beijing -- ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
