Re: [Histonet] RE: paraffin sections: white with airbubbles
Thomas- This happens when dehydration is not proper, it seems you have directly gone from 70% alcohol to paraffin, even sometimes we have faced this kind of problem in our lab even after following the right protocol. My advice is not to spend too much of time on those tissues and if u have another piece of fixed tissue , process it with standard paraffin processing protocol. Dr. Amita Dubey PCSED,TRC From: John Kiernan jkier...@uwo.ca To: Thomas, Nancy n...@stowers.org Cc: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu, 'An Eerdekens' an.eerdek...@med.kuleuven.be Date: 02/02/11 12:28 PM Subject: Re: [Histonet] RE: paraffin sections: white with airbubbles Sent by: histonet-boun...@lists.utsouthwestern.edu You can't go from 70% alcohol into paraffin without passing through 100% alcohol and then a clearing agent (liquid miscible with 100% alcohol and with melted wax). Xylene is a commonly used clearing agent. Your times in 50% and 70% alcohol are much longer than necessary. Even for a whale's hypothalamus a few hours in each solvent step should be adequate. In Belgium you may be able to obtain a great classic of histotechnology: Gabe, M (1968) Techniques histologiques. Masson et Cie, Paris. 1113 pages! This explains tissue processing very thoroughly. The author was an academic zoologist who did all his own lab work and made sure he knew what he was about. Get a copy if you can. The English translation of Manfred Gabe's book was posthumously published in 1976. I bought one then and learned a lot from it. Like many histotechnology classics, this book is now a prize possession, almost unobtainable on web sites for second-hand books. 'nuff sed. John Kiernan UWO = = = -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet- boun...@lists.utsouthwestern.edu] On Behalf Of An Eerdekens Sent: Friday, January 28, 2011 4:18 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] paraffin sections: white with airbubbles Dear collegues, I am experiencing following problem. I have embedded hypothalamus tissue in paraffin using the following procedure: -fixation in 4% paraformaldehyde for 48 hours, fixation of the tissue in 50% alcohol, next day in 70% alcohol,next day paraffin embedding. During the paraffin embedding there was a short circuit and the machine did not work for any hours, so there was a delay in the process. Now I am making slices of 5 micrometer, using the Microm HM 360. The tissue is very white (looks like I am making much thicker sections) on the slices with airbells inside. I don't have an explanation for this and many samples are showing the same features. Does someone know what might be the reason? Thanks for the help. Regards, An Eerdekens Laboratory of Intensive Care Medicine Catholic University Leuven, Belgium 003216330518 ___ 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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] stains for visualizing new bone growth
Robin, I typically stain first with Von Kossa and then counter with MacNeal's. This provides a very nice contrast where obviously mature mineralized bone is black and newly formed bone (osteoid) is grayish-green color. Additionally, your marrow space is nicely contrasted with clear visualization of osteoblasts and osteoclasts lining the bone surface. At the microscope this is a one-stop-shop stain for collecting static bone histomorphometry. Another nice contrasting stain is a modified Goldner's trichrome stain. With this stain cell nuclei are stained first with a Weigert's (iron) hematoxylin, then newly formed bone (osteoid) is stained red with an acid fuchsin/ponceau stain, next an orange G cytoplasmic stain covers the rest and a light green SF yellowish stain follows up with a nice green contrast of the mineralized bone. Very clear differentiation between mineralized bone (green) and newly formed bone (red). This stains works very well with auto threshold functions on some histomorph systems as it has a very nice contrast for the software to recognize. Of course Masson's trichrome works as well but it is typically used on decalcified paraffin embedded sections. I have found the Masson's staining kit at Sigma-Aldrich and kits for all the other stains mentioned can be found at Dorn and Hart Microedge. In fact, Dorn and Hart Microedge (www.dornandhart.com) has a lot to offer now with regards to mineralized bone (hard tissue with or without implant materials) and resin embedded histology. Good luck to you and let me know if you have any additional questions. I would also be happy to share images with you if interested. Jack On Feb 1, 2011, at 1:44 PM, Robin Dean robin_d...@compbio.com wrote: Does anyone know of a good stain to use to clearly show new bone growth other than von Kossa stain? Would appreciate any suggestions anyone might have. Thank you, Robin Robin R. Dean, Ph.D. Senior Scientist Study Director Comparative Biosciences, Inc. 786 Lucerne Dr. Sunnyvale, CA (408) 738-8060 robin_d...@compbio.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
Re: [Histonet] Off Topic: researchers very funny
This is hilarious. I got it on facebook yesterday and sent it to everyone. Paula K. Pierce, AAS, BA, HTL(ASCP)HT President Excalibur Pathology, Inc. 631 N Broadway Moore, OK 73160 405-759-3953 Lab 405-759-7513 Fax www.excaliburpathology.com From: Kimberly Tuttle ktut...@umm.edu To: histonet@lists.utsouthwestern.edu Sent: Tue, February 1, 2011 9:29:25 PM Subject: [Histonet] Off Topic: researchers very funny If this is a repost I apologize :) http://www.youtube.com/watch?v=Fl4L4M8m4d0feature=player_embedded This e-mail and any accompanying attachments may be privileged, confidential, contain protected health information about an identified patient or be otherwise protected from disclosure. State and federal law protect the confidentiality of this information. If the reader of this message is not the intended recipient; you are prohibited from using, disclosing, reproducing or distributing this information; you should immediately notify the sender by telephone or e-mail and delete this e-mail. ___ 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] Eosin stain for determining new bone old bone
Robin this is the protocol we use to showing new bone old bone. Procedure for Eosin Y 0.6 % For Staining Bone Specimen Chemicals EosinY, Sigma E-4382, 100% Ethanol, Phloxine B (Sigma P-4030) Orange G(sodium salt-Sigma O-1625) Preparation: Stock 0.6% Eosin Add 6g of eosin to 900ml 100% Ethanol and100 ml of H20; stir to dissolve. Add 50 ml glacial acetic acid adjust PH to 4.6 and 5.0. The color of the solution will change from opaque green to clear red. Stock 1% Phloxine B Solution: Dissolve 1g of Phloxine B in 100 ml distilled water and filter to make a 1% solution. Stock 2% Orange G Solution: Dissolve 2 g of Orange G in 100ml distilled water and filter to make a 2% solution. Working Solution Eosin 0.6% Make a working solution by adding 6 ml of 1% Phloxine and 6 ml of 2% Orange G to 238 ml of 0.6% eosin. Old bone dark orange. New bone light orange. Stain 15 to 30 seconds depending on what intensity you prefer. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] TTF-1 Antibody
Good Morning, we are using TTF-1 antibody from Cell Marque (clone 8G7G3/1 mouse monoclonal) on a DAKO autostainer with an LSAB+ platform. We do heat retrieval in the DAKO pascal pressure cooker using Cell Marques Trilogy retrieval solution. We do a lower temp for a longer period of time during the retrieval. I incubate the primary antibody for 1 hour and I still have problems getting the marker to work with any consistency. Where do the rest of you doing this marker get your antibody from and do you have problems getting it to work consistently? Any and all feedback would be appreciated, I'm getting really frustrated. Thanks Joanne Clark, HT Histology Supervisor Pathology Consultants of New Mexico ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] TTF-1 Antibody
We purchase out TTF-1 antibody from Ventana Medical and use it on their Benchmark XT stainer. I do believe they might purchase this from Cell Marque but I could be wrong. Anyway, we have no problem with this stain. Beatrice Sullivan, HT(A.S.C.P.) HTL , AAS, CLSP(N.C.A.) AP Supervisor Shore Memorial Hospital 609-653-3590 Speak only well of people and you need never whisper Joanne Clark jcl...@pcnm.com Sent by: To histonet-bounces@ histonet@lists.utsouthwestern.edu lists.utsouthwest cc ern.edu Subject [Histonet] TTF-1 Antibody 02/02/2011 11:18 AM Good Morning, we are using TTF-1 antibody from Cell Marque (clone 8G7G3/1 mouse monoclonal) on a DAKO autostainer with an LSAB+ platform. We do heat retrieval in the DAKO pascal pressure cooker using Cell Marques Trilogy retrieval solution. We do a lower temp for a longer period of time during the retrieval. I incubate the primary antibody for 1 hour and I still have problems getting the marker to work with any consistency. Where do the rest of you doing this marker get your antibody from and do you have problems getting it to work consistently? Any and all feedback would be appreciated, I'm getting really frustrated. Thanks Joanne Clark, HT Histology Supervisor Pathology Consultants of New Mexico ___ 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] Her2 Fixation Requirement
Hello histoland I was just given the task to find a solution that is easy and will also comply with the CAP guideline for formalin fixation documentation, and so I started my research on the Histonet archives. I found some good information, but was hoping to get more feedback. Would you mind sharing with me the actions you are taking to comply with the guideline? We are a private lab and we provide histology/pathology service for 2 hospitals and a few surgery centers. We send our blocks to Genzyme for Her2, and we must document on their test order sheet how many hours the tissues have been fixed in formalin. I'm assuming I will need to start keeping a log here, with documentation that shows what time the tissue was excised and placed in formalin from the OR, and then documentation that shows the time it was dissected and then placed in the tissue processor. The problem that I may come across is getting the OR nurse to document the time for us. I don't know...maybe we need to put another sections on our requisition form, or maybe something on the formalin container itself for the nurse to write on. It'll be a hassle at first but if I can get the hospitals lab director involved, I'm sure it will work itself out. Anyway, if you wouldn't mind sharing some of your ideas, I would really appreciate it. Paula Lucas Lab Manager Bio-Path Medical Group Fountain Valley, CA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Her2 Fixation Requirement
We receive our lumpectomies and total breasts fresh. They are immediately put in 10 % formalin if no frozen is required and that time is noted on our requisition. Our processing time for formalin and the fixation time is made part of the final report. If we receive a breast biopsy specimen we have asked that they write the time on the requistion that the specimen was placed in 10 % formalin. We set up a procedure that states this. Beatrice Sullivan, HT(A.S.C.P.) HTL , AAS, CLSP(N.C.A.) AP Supervisor Shore Memorial Hospital 609-653-3590 Speak only well of people and you need never whisper Paula Lucas plucas@biopath.o rgTo Sent by: histonet@lists.utsouthwestern.edu histonet-bounces@ cc lists.utsouthwest ern.edu Subject [Histonet] Her2 Fixation Requirement 02/02/2011 12:11 PM Hello histoland I was just given the task to find a solution that is easy and will also comply with the CAP guideline for formalin fixation documentation, and so I started my research on the Histonet archives. I found some good information, but was hoping to get more feedback. Would you mind sharing with me the actions you are taking to comply with the guideline? We are a private lab and we provide histology/pathology service for 2 hospitals and a few surgery centers. We send our blocks to Genzyme for Her2, and we must document on their test order sheet how many hours the tissues have been fixed in formalin. I'm assuming I will need to start keeping a log here, with documentation that shows what time the tissue was excised and placed in formalin from the OR, and then documentation that shows the time it was dissected and then placed in the tissue processor. The problem that I may come across is getting the OR nurse to document the time for us. I don't know...maybe we need to put another sections on our requisition form, or maybe something on the formalin container itself for the nurse to write on. It'll be a hassle at first but if I can get the hospitals lab director involved, I'm sure it will work itself out. Anyway, if you wouldn't mind sharing some of your ideas, I would really appreciate it. Paula Lucas Lab Manager Bio-Path Medical Group Fountain Valley, CA ___ 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] Her2 Fixation Requirement
We have the nurse document the time of removal on the requisition. The next requisition update will have a spot for this. They have been very cooperative and do a good job. In addition, you must document the cold ischmic time - that is the time from removal until time in formalin. This is important when the specimen goes for xray or whatever. So there is a removal time, an into formalin time and an out of formalin time. Then if we don't have time allowed to meet the fixation time with the regular, it is put on a late processor, if we have one available, or it is held overnight. And if it has to come off on Sunday, we have a med tech remove it from the processor and it waits for us to come on Monday to embed it. The pathologists document all this time in the report. Best! J Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 678-843-7376 - Phone 678-843-7831 - Fax -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Paula Lucas Sent: Wednesday, February 02, 2011 12:11 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Her2 Fixation Requirement Hello histoland I was just given the task to find a solution that is easy and will also comply with the CAP guideline for formalin fixation documentation, and so I started my research on the Histonet archives. I found some good information, but was hoping to get more feedback. Would you mind sharing with me the actions you are taking to comply with the guideline? We are a private lab and we provide histology/pathology service for 2 hospitals and a few surgery centers. We send our blocks to Genzyme for Her2, and we must document on their test order sheet how many hours the tissues have been fixed in formalin. I'm assuming I will need to start keeping a log here, with documentation that shows what time the tissue was excised and placed in formalin from the OR, and then documentation that shows the time it was dissected and then placed in the tissue processor. The problem that I may come across is getting the OR nurse to document the time for us. I don't know...maybe we need to put another sections on our requisition form, or maybe something on the formalin container itself for the nurse to write on. It'll be a hassle at first but if I can get the hospitals lab director involved, I'm sure it will work itself out. Anyway, if you wouldn't mind sharing some of your ideas, I would really appreciate it. Paula Lucas Lab Manager Bio-Path Medical Group Fountain Valley, CA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: This e-mail, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please delete this message, and reply to the sender regarding the error in a separate email. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Looking for Part time work in Phoenix AZ
Hi all, I am looking for part time work in the Phoenix area, I am HT ascp and have 18 years experience. Please respond to this email, thank you. Jill ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Rat lung histology for mast cells and eosinophils
Chen For mast cells you can fix in 10% NBF and be fine with toluidine blue. We stain with toluidine blue on formalin fixed samples all of the time. The trick is not to dehydrate the sections, let them air dry and then mount. I would stick with formalin fixation that's going to be your best bet for most specials and IHC stains. For eosinophils you can stain with giemsa, or we would run a modified dif-quik stain that worked well for eosinophils. For neutrophils I'm not sure but if you are staining for mouse neutrophils, serotec has a nice antibody. Macrophages would be IHC for ED-1 also from serotec, CD3 from Dako will work on rat tissue nicely. There are other markers that will work on cell lines. Good Luck Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Chen, Shu-Cheng Sent: Wednesday, February 02, 2011 11:29 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Rat lung histology for mast cells and eosinophils Hi, We are called to support an asthmatic rat lung project and need to stain for mast cells, eosinophils, neutrophils and possibly other inflammatory cells, such as Th2 cells etc. Any suggestions you can give us in terms of fixatives and special stains or IHC are very much appreciated. From my search, it seems that Carnoy's fixative with toluidine blue is the way to go for mast cells. But can one clearly differentiate eos from neuts with this fixative or a formalin fixed HE stained lung section? If we need to do Trichrome, PAS, May Grunwald Giemsa and HE etc. what would be the best fixative to accommodate these stains? BTW, I heard that formalin free Zn-fixative is good for IHC. Will it be good for all these stains above? Sorry for so many questions. It shows how little experience I have in this area. Thank you, Shu 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] re Her2 neu requirements
We have the specimens sent fresh to our lab and we put them in formalin there. We document the time and the we make sure all are processed in the time window. The diffculty is usually on Friday biopsy and excision specimens, especially if you have a courier run. John Connelly, M.D. Hello histoland I was just given the task to find a solution that is easy and will also comply with the CAP guideline for formalin fixation documentation, and so I started my research on the Histonet archives. I found some good information, but was hoping to get more feedback. Would you mind sharing with me the actions you are taking to comply with the guideline? We are a private lab and we provide histology/pathology service for 2 hospitals and a few surgery centers. We send our blocks to Genzyme for Her2, and we must document on their test order sheet how many hours the tissues have been fixed in formalin. I'm assuming I will need to start keeping a log here, with documentation that shows what time the tissue was excised and placed in formalin from the OR, and then documentation that shows the time it was dissected and then placed in the tissue processor. +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
RE: [Histonet] Her2 Fixation Requirement
What is the appropriate time it should be in formalin? Do you start your time when it is initially put into formalin, or the start from the time it was dissected? Lori A Disher Fawcett Memorial Hospital Port Charlotte, FL 33952 lori.dis...@hcahealthcare.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Histonet Digest, Vol 87, Issue 4
We too are a small lab in California, and faced these same questions. We simply document all this in the report. For example; we know that our tissue comes out of formalin on the processor at 2330. We have the surgery staff document the time the specimen went into formalin. We then subtract that time form 2330 and have the hours in formalin. You are correct that it took a great deal of effort to see that surgery complied, but after they realized we were relentless with phone calls, things fell into place. Oh yes! On the weekend someone must come in to remove the tissue from the processor. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of histonet-requ...@lists.utsouthwestern.edu Sent: Wednesday, February 02, 2011 10:01 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 87, Issue 4 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. Re: Off Topic: researchers very funny (Paula Pierce) 2. Eosin stain for determining new bone old bone (Jacquitta Taylor) 3. TTF-1 Antibody (Joanne Clark) 4. Re: TTF-1 Antibody (bsulli...@shorememorial.org) 5. Her2 Fixation Requirement (Paula Lucas) 6. Re: Her2 Fixation Requirement (bsulli...@shorememorial.org) 7. RE: Her2 Fixation Requirement (Weems, Joyce) -- Message: 1 Date: Wed, 2 Feb 2011 05:49:57 -0800 (PST) From: Paula Pierce cont...@excaliburpathology.com Subject: Re: [Histonet] Off Topic: researchers very funny To: Kimberly Tuttle ktut...@umm.edu, Histonet histonet@lists.utsouthwestern.edu Message-ID: 217328.82615...@web1112.biz.mail.sk1.yahoo.com Content-Type: text/plain; charset=iso-8859-1 This is hilarious. I got it on facebook yesterday and sent it to everyone. Paula K. Pierce, AAS, BA, HTL(ASCP)HT President Excalibur Pathology, Inc. 631 N Broadway Moore, OK 73160 405-759-3953 Lab 405-759-7513 Fax www.excaliburpathology.com From: Kimberly Tuttle ktut...@umm.edu To: histonet@lists.utsouthwestern.edu Sent: Tue, February 1, 2011 9:29:25 PM Subject: [Histonet] Off Topic: researchers very funny If this is a repost I apologize :) http://www.youtube.com/watch?v=Fl4L4M8m4d0feature=player_embedded This e-mail and any accompanying attachments may be privileged, confidential, contain protected health information about an identified patient or be otherwise protected from disclosure. State and federal law protect the confidentiality of this information. If the reader of this message is not the intended recipient; you are prohibited from using, disclosing, reproducing or distributing this information; you should immediately notify the sender by telephone or e-mail and delete this e-mail. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Message: 2 Date: Wed, 2 Feb 2011 09:13:40 -0600 From: Jacquitta Taylor jacqui...@earthlink.net Subject: [Histonet] Eosin stain for determining new bone old bone To: histonet@lists.utsouthwestern.edu Message-ID: 4c4b1f98-d6ef-45df-a010-851cffe11...@earthlink.net Content-Type: text/plain; charset=us-ascii Robin this is the protocol we use to showing new bone old bone. Procedure for Eosin Y 0.6 % For Staining Bone Specimen Chemicals EosinY, Sigma E-4382, 100% Ethanol, Phloxine B (Sigma P-4030) Orange G(sodium salt-Sigma O-1625) Preparation: Stock 0.6% Eosin Add 6g of eosin to 900ml 100% Ethanol and100 ml of H20; stir to dissolve. Add 50 ml glacial acetic acid adjust PH to 4.6 and 5.0. The color of the solution will change from opaque green to clear red. Stock 1% Phloxine B Solution: Dissolve 1g of Phloxine B in 100 ml distilled water and filter to make a 1% solution. Stock 2% Orange G Solution: Dissolve 2 g of Orange G in 100ml distilled water and filter to make a 2% solution. Working Solution Eosin 0.6% Make a working solution by adding 6 ml of 1% Phloxine and 6 ml of 2% Orange G to 238 ml of 0.6% eosin. Old bone dark orange. New bone light orange. Stain 15 to 30 seconds depending on what intensity you prefer. -- Message: 3 Date: Wed, 2 Feb 2011 09:18:23 -0700 From: Joanne Clark jcl...@pcnm.com Subject: [Histonet] TTF-1 Antibody To: histonet@lists.utsouthwestern.edu Message-ID:
RE: [Histonet] Her2 Fixation Requirement
Lori I would think both if you could since the Her2 paper does address those issues as potentials for problems in testing variation (time to fixation). I believe the CAP/ASCO paper states not less then 6 and no more than 48 hours for fixation (2007 paper). I believe you also should document type of fixative if possible; vendor, lot number and expiration date if you can. I have pdfs of these documents if you need them. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of lori.dis...@hcahealthcare.com Sent: Wednesday, February 02, 2011 12:16 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Her2 Fixation Requirement What is the appropriate time it should be in formalin? Do you start your time when it is initially put into formalin, or the start from the time it was dissected? Lori A Disher Fawcett Memorial Hospital Port Charlotte, FL 33952 lori.dis...@hcahealthcare.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
RE: RE: [Histonet] Her2 Fixation Requirement
Formalin time is no less than 6 and no more than 48 hours. You need to document time from excision to time in formalin - cold ischemic time - and time in formalin. New regs extended fixation time for ER/PR to 72 hours, but since it's all in there together, you can't go by that yet. And for patient's to be eligible for many clinical trials, 10% NBF is the only fixative that should be used. j -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of lori.dis...@hcahealthcare.com Sent: Wednesday, February 02, 2011 14:16 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Her2 Fixation Requirement What is the appropriate time it should be in formalin? Do you start your time when it is initially put into formalin, or the start from the time it was dissected? Lori A Disher Fawcett Memorial Hospital Port Charlotte, FL 33952 lori.dis...@hcahealthcare.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: This e-mail, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please delete this message, and reply to the sender regarding the error in a separate email. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Her2 Fixation Requirement
I believe the two most recent articles from ASCO/CAP on this subject are Hammond, M.E., D.F. Hayes, M. Dowsett, D.C. Allred, K.L. Hagerty, S. Badve, P.L. Fitzgibbons, G. Francis, et al. 2010. American Society of Clinical Oncology/College Of American Pathologists guideline recommendations for immunohistochemical testing of estrogen and progesterone receptors in breast cancer. J Clin Oncol 28:2784-2795. Wolff, A.C., M.E. Hammond, J.N. Schwartz, K.L. Hagerty, D.C. Allred, R.J. Cote, M. Dowsett, P.L. Fitzgibbons, et al. 2007. American Society of Clinical Oncology/College of American Pathologists guideline recommendations for human epidermal growth factor receptor 2 testing in breast cancer. J Clin Oncol 25:118-145. If you do not have access to the literature you can request reprints from the authors or it may be available on the ASCO website? Luis -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Liz Chlipala Sent: Wednesday, February 02, 2011 2:28 PM To: lori.dis...@hcahealthcare.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Her2 Fixation Requirement Lori I would think both if you could since the Her2 paper does address those issues as potentials for problems in testing variation (time to fixation). I believe the CAP/ASCO paper states not less then 6 and no more than 48 hours for fixation (2007 paper). I believe you also should document type of fixative if possible; vendor, lot number and expiration date if you can. I have pdfs of these documents if you need them. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of lori.dis...@hcahealthcare.com Sent: Wednesday, February 02, 2011 12:16 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Her2 Fixation Requirement What is the appropriate time it should be in formalin? Do you start your time when it is initially put into formalin, or the start from the time it was dissected? Lori A Disher Fawcett Memorial Hospital Port Charlotte, FL 33952 lori.dis...@hcahealthcare.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 email message, including any attachments, is for the sole use of the intended recipient(s) and may contain information that is proprietary, confidential, and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure, or distribution is prohibited. If you have received this email in error please notify the sender by return email and delete the original message. Please note, the recipient should check this email and any attachments for the presence of viruses. The organization accepts no liability for any damage caused by any virus transmitted by this email. = ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] WindoPath Users
How many folks are using this LIS for AP? Any feedback would be appreciated. Thanks! Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 678-843-7376 - Phone 678-843-7831 - Fax Confidentiality Notice: This e-mail, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please delete this message, and reply to the sender regarding the error in a separate email. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Trace Gas companies
Hi Histonetters Can you all share with me where you get and send in your trace gas analysis badges? Thanks a bunch! :-D Kim Donadio Pathology Supervisor Baptist Hospital 1000 W Moreno St. Pensacola FL 32501 Phone (850) 469-7718 Fax (850) 434-4996 - All electronic data transmissions originating from or sent to Baptist Health Care Corporation (BHC) are subject to monitoring. This message along with any attached data, are the confidential and proprietary communications of BHC and are intended to be received only by the individual or individuals to whom the message has been addressed. If the reader of this message is not the intended recipient, please take notice that any use, copying, printing, forwarding or distribution of this message, in any form, is strictly prohibited and may violate State or Federal Law. If you have received this transmission in error, please delete or destroy all copies of this message. For questions, contact the BHC Privacy Officer at (850) 434-4472. Rev.10/07. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] WindoPath Users
I have been using WindoPath for a year and a half now and I love it! We are a GI lab so building our library was not as time consuming as a it would be for a full spectrum path lab, but you only have to do that once. We have had four pathologists trained on using the program and they all find it very user friendly as well. Feel free to contact me should you like more information. Thanks, Cristi Sent from my HTC on the Now Network from Sprint! - Reply message - From: Weems, Joyce jwe...@sjha.org Date: Wed, Feb 2, 2011 12:18 pm Subject: [Histonet] WindoPath Users To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu How many folks are using this LIS for AP? Any feedback would be appreciated. Thanks! Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 678-843-7376 - Phone 678-843-7831 - Fax Confidentiality Notice: This e-mail, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please delete this message, and reply to the sender regarding the error in a separate email. ___ 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] Safranin O
Hello everyone, I am trying to stain for cartilage in mouse skull (cross-sectioning with the brain in tact) and wanted to know if anyone might have a good protocol for Safranin O / Fast green or Safranin O / von Kossa? The tissue is embedded in MMA and was cut on a saw microtome and finished down to an approximate 20 to 30 micron tissue thickness - 50 microns including the glue layer (I glue a slide to the block before making the cut and then grind and polish to finish). I tried Weigert's hematoxylin / Safranin O / Fast Green ( I began with Weigert's Hematoxylin, followed by Fast Green and finished with Safranin O), but the Safranin O seemed to overpower the Fast Green (the calcified bone stained more red than green). Any help on this issue would be greatly appreciated as always. Thanks again. Jim ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] EGF IHC
Hello everyone Does anyone out there have a good EGF (not EGFR) antibody that works well in formalin fixed paraffin embedded samples - species is human Thanks Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] slide labeler advice
Hello again We are in the market for a slide labeler. We currently have the SlideMate from Thermo Scientific. We have had it for about 2 years now and we are just not happy with the quality and consistency of the printing. We are a contract research lab and not a clinical lab. Entering data for each slide is not an option. The programming that comes with the Slide Mate is nice, except that no one has really spent the time to train us on it, we have figured it out a bit. We would want a labeler that we could create formats for different sample types, so all we would have to do is put the animal ID in and the printer would print all of the slides needed. Would like barcoding capabilities just incase we move to that in the future. Any advice or suggestions are appreciated. Vendors welcome also. Thanks in advance. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] slide labeler advice
Hi Liz We love our IPS and IPC printers from leica. Both strong work horses that are adaptable to your needs. Stacey Cu Dermpath Sent from myTouch 4G - Reply message - From: Liz Chlipala l...@premierlab.com To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Subject: [Histonet] slide labeler advice Date: Wed, Feb 2, 2011 3:58 pm Hello again We are in the market for a slide labeler. We currently have the SlideMate from Thermo Scientific. We have had it for about 2 years now and we are just not happy with the quality and consistency of the printing. We are a contract research lab and not a clinical lab. Entering data for each slide is not an option. The programming that comes with the Slide Mate is nice, except that no one has really spent the time to train us on it, we have figured it out a bit. We would want a labeler that we could create formats for different sample types, so all we would have to do is put the animal ID in and the printer would print all of the slides needed. Would like barcoding capabilities just incase we move to that in the future. Any advice or suggestions are appreciated. Vendors welcome also. Thanks in advance. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.comhttp://www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504 ___ 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] CSF for cytology
Hi Histonet, We are going to be receiving some cerebrospinal fluid as party of an MS Panel for cytology. Our pathologist would like me to make a cell button and stain it with HE and Diff Quik, but I wanted to check with you guys and see if there are any other stains that I might do with this case. Thank you for your help, Katelin Lester, HTL(ASCP) MedSurg Pathology Associates, Inc. (503)443-2157 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] CSF for cytology
Katelin, CSF specimens usually contain very few cells, maybe a lymphocyte or two. Making a cell button (?cell block for Histo processing) would give disappointing results. I would recommend cytocentrifugation (at least one air-dried DQ stained and one 95% ethanol-fixed PAP stained). You could even concentrate the cells using centrifugation prior to cytocentrifugation. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Katelin Lester Sent: Thursday, 3 February 2011 12:54 PM To: histonet Subject: [Histonet] CSF for cytology Hi Histonet, We are going to be receiving some cerebrospinal fluid as party of an MS Panel for cytology. Our pathologist would like me to make a cell button and stain it with HE and Diff Quik, but I wanted to check with you guys and see if there are any other stains that I might do with this case. Thank you for your help, Katelin Lester, HTL(ASCP) MedSurg Pathology Associates, Inc. (503)443-2157 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet * This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. * ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Trace Gas companies
We just did ours this week! We get them from Environmental Monitoring Technology. http://www.emt-online.com/ Katelin Lester, HTL(ASCP) MedSurg Pathology Associates, Inc. (503)443-2157 Hi Histonetters Can you all share with me where you get and send in your trace gas analysis badges? Thanks a bunch! :-D Kim Donadio Pathology Supervisor Baptist Hospital 1000 W Moreno St. Pensacola FL 32501 Phone (850) 469-7718 Fax (850) 434-4996 - All electronic data transmissions originating from or sent to Baptist Health Care Corporation (BHC) are subject to monitoring. This message along with any attached data, are the confidential and proprietary communications of BHC and are intended to be received only by the individual or individuals to whom the message has been addressed. If the reader of this message is not the intended recipient, please take notice that any use, copying, printing, forwarding or distribution of this message, in any form, is strictly prohibited and may violate State or Federal Law. If you have received this transmission in error, please delete or destroy all copies of this message. For questions, contact the BHC Privacy Officer at (850) 434-4472. Rev.10/07. ___ 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] slide labeler advice
Hi Liz, As a provider of on-demand-printing solutions I've worked closely with labs using Thermo SlideMate printers. I've noticed a couple of simple maintenace steps that may dramatically improve your printing results. First, clean the print head daily. To do this, press the Setup button, then press Load. This releases the takeup spool that keeps pressure on the ribbon. Now, open the door and loosen the ribbon where it wraps around the print head - that's the half gold/half silver rectangular bar and the print head is at the lowest point where it comes to a V shape. Wet a Kwik Wipe with alcohol and gently clean the print head. If you haven't done this in a while you'll want to clean it well the first time around. Get the Kwik Wipe good and wet (but not dripping) and dab the alcohol on, let it sit for a few seconds, then wipe dry. Do it a few times using a new clean wipe and get all around the V shape. Also wipe away any dust or dirt around the inside. After that initial cleaning, you should only need to do a quick wipe it clean, but make it a part of daily maintenance at the start or end of each day. Before you close the door, wind the takeup spool to remove any slack in the tape. Second, remove excess tape from the takeup spool. As more tape winds up on the takeup spool the tension can become spongy. This can cause the tape to crease which will leave a white line in the printed area. When you do your daily maintenance, just strip the day's tape from the takeup spool and re-thread it. Performing these steps daily should take 2 - 3 minutes and should keep print quality consistent. There are other things I've come across for programming and utilities for the SlideMate, etc. but too much to include here. Also, we provide our own print-on-demand software solution that works with the SlideMate and other label printers. Feel free to post additional questions, or contact me off line. Best Regards, Brian Brian Wood Detangle IT, Inc. 516 594-9344 br...@detangleit.commailto:br...@detangleit.com On Feb 2, 2011, at 5:56 PM, Liz Chlipala l...@premierlab.commailto:l...@premierlab.com wrote: Hello again We are in the market for a slide labeler. We currently have the SlideMate from Thermo Scientific. We have had it for about 2 years now and we are just not happy with the quality and consistency of the printing. We are a contract research lab and not a clinical lab. Entering data for each slide is not an option. The programming that comes with the Slide Mate is nice, except that no one has really spent the time to train us on it, we have figured it out a bit. We would want a labeler that we could create formats for different sample types, so all we would have to do is put the animal ID in and the printer would print all of the slides needed. Would like barcoding capabilities just incase we move to that in the future. Any advice or suggestions are appreciated. Vendors welcome also. Thanks in advance. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 http://www.premierlab.comwww.premierlab.comhttp://www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504 ___ Histonet mailing list mailto:Histonet@lists.utsouthwestern.eduHistonet@lists.utsouthwestern.edumailto:Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonethttp://lists.utsouthwestern.edu/mailman/listinfo/histonet This message scanned by Reflexion Total Control___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Safranin O
Have you tried Sanderson's Rapid Bone Stain? Try using it first (7 minutes at 60C), then rinse in dH2O (a few dip an dunks at 60C and blot dry), then counterstain with safranin O for 2-5 minutes (room temp - check intensity) and rinse in 100% EtOH (room temp - few dip and dunks and blot dry). If saf o is too intense, you should be able to differentiate a little with 95% EtOH first then do the 100% EtOH step. On a curious note, is there any particular reason why you are cutting thick sections and grinding? You should very easily be able to cut thin sections with a rotary microtome using a tungsten-carbide knife at 5 microns and then you would have greater staining flexibility. Call me if you want to discuss (317-281-1975). Jack On Feb 2, 2011, at 4:18 PM, Herrick, James L. (Jim) herrick.ja...@mayo.edu wrote: Hello everyone, I am trying to stain for cartilage in mouse skull (cross-sectioning with the brain in tact) and wanted to know if anyone might have a good protocol for Safranin O / Fast green or Safranin O / von Kossa? The tissue is embedded in MMA and was cut on a saw microtome and finished down to an approximate 20 to 30 micron tissue thickness - 50 microns including the glue layer (I glue a slide to the block before making the cut and then grind and polish to finish). I tried Weigert's hematoxylin / Safranin O / Fast Green ( I began with Weigert's Hematoxylin, followed by Fast Green and finished with Safranin O), but the Safranin O seemed to overpower the Fast Green (the calcified bone stained more red than green). Any help on this issue would be greatly appreciated as always. Thanks again. Jim ___ 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
