Re: [Histonet] Processing Fat for Paraffin
Paula: For me the times (2 h/each), your dependable instrument and the schedule seem to be OK, but I am not so sure about the Citrisolve because this product (1.4 more expensive than xylene) is made of 91% of water and surfactants + 9% d-limonene and I am not very confident that such a clearing agent will be able to allow a proper infiltration on your specimens. At least for these specimens I would use xylene or a mixture of 2-propanol and mineral oil (attached to another e-mail to you). René J. --- On Thu, 4/30/09, Va Paula Sicurello vapat...@yahoo.com wrote: From: Va Paula Sicurello vapat...@yahoo.com Subject: [Histonet] Processing Fat for Paraffin To: HistoNet histonet@lists.utsouthwestern.edu, MSA BB microsc...@microscopy.com Date: Thursday, April 30, 2009, 6:22 PM Hello out there in fat processing land, I have been given human fat samples and need to embed them in paraffin. In the past I've used a VIP processor for this and now I have an Autotechnicon (vintage dual model) with a timing wheel. I know I need to process these fatties slowly, my question is--can I use 2 hours per step and have it turn out OK? I have a timing wheel punched out for 2 hour steps. My steps would be alcohols: 70, 80, 95 x 2, 100 x 3, Citrisolv x3, paraffin x2 and another paraffin step under vacuum. Let me know your wise and experienced opinions or protocols. I don't have anything else to use except the 43 year old Autotechnicon so don't even suggest it. You make me feel bad that I can't get my research foundation to buy me something new or even newer. ;-) Stewing in somebody else's fat (eew!), Paula :-) Paula Sicurello VA Medical Center San Diego Veterans Medical Research Foundation (VMRF) Core Research Imaging Center 3350 La Jolla Village Dr., MC151 San Diego, CA 92161 858-552-8585 x2397 ___ 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] Reuse of citrate buffer
Just went to the TriState Meeting in Madison Wisconsin and attended a very good lecture on immunohistochemistry focusing on antigen retrieval and it was stressed that one should not reuse any antigen retrieval solution. It should be made fresh when needed. Dolores The information transmitted is intended only for the person(s)or entity to which it is addressed and may contain confidential and/or legally privileged material. Delivery of this message to any person other than the intended recipient(s) is not intended in any way to waive privilege or confidentiality. Any review, retransmission, dissemination or other use of , or taking of any action in reliance upon, this information by entities other than the intended recipient is prohibited. If you receive this in error, please contact the sender and delete the material from any computer. For Translation: http://www.baxter.com/email_disclaimer ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Credentials for performing IHC staining
Wew are having an ongoing debate as to the required credentials for a person to perform IHC staining on the Ventana instruments. Can you help ? Thanks. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] OCT and RNA
Hi, Some of our researchers want to collect frozen tissue for possible future use for frozen sections or for RNA analysis. We would like to know how freezing in OCT will effect subsequent RNA determinations if it is decided that they need the tissue more for RNA analysis than for frozen sections for morphology or IHC studies. Thanks very much for you adivice. Karen _ Hotmail® has a new way to see what's up with your friends. http://windowslive.com/Tutorial/Hotmail/WhatsNew?ocid=TXT_TAGLM_WL_HM_Tutorial_WhatsNew1_052009___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Credentials for performing IHC staining
Regardless of the instrument, or if IHC is done manually, it is considered a high complexity test and the person doing it should be an accredited or licensed, specially trained, histotechnologist or med tech specialized in IHC which is a new ASCP accreditation level. René J. --- On Fri, 5/1/09, jimr0...@comcast.net jimr0...@comcast.net wrote: From: jimr0...@comcast.net jimr0...@comcast.net Subject: [Histonet] Credentials for performing IHC staining To: Histonet histonet-requ...@lists.utsouthwestern.edu, Histonet2 histonet@lists.utsouthwestern.edu Date: Friday, May 1, 2009, 10:18 AM Wew are having an ongoing debate as to the required credentials for a person to perform IHC staining on the Ventana instruments. Can you help ? 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
[Histonet] Texas Society for Histotechnology 2009
Hello Was interested in receiving cost information on the meeting in Austin. Would be driving down on Saturday morning, for Saturday attendance only. Possibly to attend one or two sessions depending on start time. Am not a member of Texas society. Am registered HTL/CT ASCP. Please send appropriate schedules, forms, and information. Thanks Jeanette Montgomery Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please destroy all copies of the original message. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Need refurbished HE stainer
Sally; I am the rep from Dako. Would you please provide your contact information? Thanks Chuck Rippin ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] xylene
We have recently had a problem with poor tissue processing (soft to cut/ poor staining). None of the tissue was processed the first time, and ultimately, some fragile tissue did not survive. The alcohols checked out and the processor, itself, does not appear to be an issue. We used recycled xylene in the processor first, followed by fresh xylene. The xylene, which came from a CBG recycler, failed the test for contamination, after the fact. Has anyone seen this kind of problem? We would appreciate any input. Thank you. Loretta Moffatt, MT(ASCP),MHA Laboratory Manager 777 Rural Avenue Williamsport, PA 17701 570-321-2326 (F)570-321-2489 lmoff...@susquehannahealth.org Patience is never more important than when you are on the verge of losing it. Confidentiality Notice: This message and any attachments originate by electronic mail from Susquehanna Health System and their subsidiaries/affiliates (SHS). Both this document and any attachments are intended for the sole use of the addressee indicated above and may contain proprietary, privileged and/or confidential information. If you are not the intended recipient of this message, you are hereby notified that any use or disclosure of this information is strictly prohibited. If you received this message in error, or have reason to believe you are not authorized to receive it, please notify the sender by reply email, with a copy to itsecur...@susquehannahealth.org mailto:itsecur...@susquehannahealth.org, and then promptly delete the original and reply messages. Thank you for your cooperation. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Frozen Tonsil Controls
Does anyone know where to purchase frozen tonsil control tissue for IF's? Thanks! Jennifer Bull Histology Supervisor jennifer.b...@nwpathology.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] OCT and RNA
I don't think the OCT will affect the RNA in the tissue, but Ambion has a product and resources that will help you confirm that and protect your RNA. Here's the link to the product: http://www.ambion.com/catalog/CatNum.php?AM7030 And on their home page they have a link to their tech support dept. which has articles from the basics of RNA on up. Hope this helps! Kathleen Roberts Principal Lab Technician Neurotoxicology Labs Dept of Pharmacology Toxicology Ernest Mario School of Pharmacy Rutgers, the State University of NJ 41 B Gordon Rd Piscataway, NJ 08854 (732) 445-6914 Karen Doty wrote: Hi, Some of our researchers want to collect frozen tissue for possible future use for frozen sections or for RNA analysis. We would like to know how freezing in OCT will effect subsequent RNA determinations if it is decided that they need the tissue more for RNA analysis than for frozen sections for morphology or IHC studies. Thanks very much for you adivice. Karen _ Hotmail® has a new way to see what's up with your friends. http://windowslive.com/Tutorial/Hotmail/WhatsNew?ocid=TXT_TAGLM_WL_HM_Tutorial_WhatsNew1_052009___ 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] FISH together with IF
Dear Histonetters, In the context of Graft-versus-Host-Disease I’m trying to couple immunfluorescence with FISH. I agree, it is a bit daring to do FISH and IF at once. The problem is that IF is suffering from the pre-treatment during FISH. After 2 failed stainings I was able to state that primary antibodies of IF couldn’t bind antigens after FISH. The question that rose concerning this over-digestion of antigens is – could I omit the pre-treatment with pepsin, but still but still get a satisfying FISH? Could you, please, look through my protocol. Maybe I’m making the same mistakes you have also encountered. With kind regards, Wladislav Sokalskiy, medical student at the University of Mainz, Germany DAY 1: 1. Putting the section in incubator for 10 min at 70 C. 2. Washing in xylol for 4 x 5 min and rehydrogenating for 5 min in Ethanol. 3. 5 min in ddH2O. 4. For antigen retrieval: washing for 20 min at ~ 100 C in Na-Citrat-Buffer (ph 6). 5. 6 min in Washing-Buffer (don’t know exactly what it consists of, because it was delivered within the FISH-kit) 6. Digestion for 2,5 min with Pepsin. 7. 6 min in Washing-Buffer 8. Dehydrogenization with Ethanol 9. Drying for 12 min at 37 C (is it may be better to dry at the room temperature) 10. Hybridizing for 10 min at 75 C and keeping the section overnight at 37 C (I haven’t coverslipped the section as I kept it overnight. Should I? Was it a mistake on my part?) DAY 2: 1. Washing the section for 5 min in TBS (with 0,05 % Tween 20) 2. Blocking the unspecific antigens with TNT (with 0,05% Tween 20) for 30 min 3. Adding primary antibody (diluted 1:100) 4. Keeping incubating overnight in the moisture chamber DAY 3: 1. Washing 2 x 15 min in TBS (with Tween) 2. Adding secondary antibody (Alexa Fluor 488 – diluted 1:300) and keeping it incubating for 2 h 3. Washing it 2 x 15 min in TBS (with Tween) 4. Adding DAPI covering the section with glass. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Frozen Tonsil Controls
I don't know where to purchase them but if you have a childrens hospital nearby they can probably accommodate you. Hazel Horn Hazel Horn, HT/HTL (ASCP) Supervisor of Histology Arkansas Children's Hospital 1 Children's WaySlot 820 Little Rock, AR 72202 phone 501.364.4240 fax501.364.3155 visit us on the web at:www.archildrens.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bull, Jennifer L. Sent: Friday, May 01, 2009 12:31 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Frozen Tonsil Controls Does anyone know where to purchase frozen tonsil control tissue for IF's? Thanks! Jennifer Bull Histology Supervisor jennifer.b...@nwpathology.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] OCT and RNA
We have taken sections from OCT embedded tissues and processed them for RNA analysis with out any problems. You should be careful to move the tissue to a solution that will protect the RNA once it has been either sectioned or thawed. Good Luck! Trudeau Institute email service will be temporarily suspended on May 2-3 to complete the joining of electrical services between the newly constructed Stafford Wing and our existing facilities. Email sent to and from all Trudeau Institute employees will be held in queue until services are restored on Monday, May 4th ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] STP420
Good Afternoon, We have the thermofisher stp 420 processor. We are having major cross contamination problems. Finding brain and placenta fragments in with other tissue. Anyone else with this processor having similar problems? Thanks, Janice ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] STP420
I have never used this tissue processor, but I don't understand how it would be the tissue processor that would be causing the contamination. Does this processor have different processing chamber? If so, does it use the same processing solutions? Is all your tissues sectioned on the same cutting board? By the same cutter? Just throwing out some general thoughts. Mike -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Janice Mitchell Sent: Friday, May 01, 2009 3:14 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] STP420 Good Afternoon, We have the thermofisher stp 420 processor. We are having major cross contamination problems. Finding brain and placenta fragments in with other tissue. Anyone else with this processor having similar problems? Thanks, Janice ___ 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)
Can anyone tell me how to use liquid nitrogen on fatty sections and also give me a source for purchasing it and a rough estimate of how much it costs. I would appreciate any help. We have been battling difficult breast specimens at our tiny little (cheap) hospital for entirely too long. Thanks, Jennifer Johnson, HTL,(ASCP) _ Rediscover Hotmail®: Get e-mail storage that grows with you. http://windowslive.com/RediscoverHotmail?ocid=TXT_TAGLM_WL_HM_Rediscover_Storage2_042009___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Cancer Diagnostics and Surgipath
This may be just a rumor, but I heard Patrick, one of the Surgipath Sales Managers, actually set up Cancer Diagnostics and was running a side business while also still working for Surgipath. And when Surgipath was sold to Leica, Leica in doing their due diligence discovered this and summilarily dismissed him and thus cut him off of supply for stealing leads and may be taking legal action. JC. From: histoman3...@live.com To: histonet@lists.utsouthwestern.edu Date: Fri, 1 May 2009 17:47:25 -0400 Subject: [Histonet] Cancer Diagnostics and Surgipath I purchased Surgipath cassettes from Cancer Diagnostics but they do not supply these any more and I cannot reach my rep. Does anyone know why? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Cancer Diagnostics and Surgipath
I purchased Surgipath cassettes from Cancer Diagnostics but they do not supply these any more and I cannot reach my rep. Does anyone know why? _ Windows Live™ SkyDrive™: Get 25 GB of free online storage. http://windowslive.com/online/skydrive?ocid=TXT_TAGLM_WL_skydrive_042009___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
