Dear Salim, I have done lots of IHC on mouse and rat tissues ie brain and trying to make correlations on viable tissue is hard enough. If you do preform the IHC it may confound your data one way or the other and leave you with more questions then answers. My recommendation would be to leave this animal out of the analysis. Autolysis can be difficult to read through with IHC in the brain. Ultimately it is up to you but if this animal shows more or less staining and skews your average in that group you will be left with the nagging question " was it the autolysis that made the difference"
My 2 cents...good luck.. Jamie E Erickson Scientist II, M.S. HTL (ASCP) Discovery Safety, Metabolism & Pharmacokinetics Abbott Laboratories From: histonet-requ...@lists.utsouthwestern.edu To: histonet@lists.utsouthwestern.edu Date: 11/29/2009 01:02 PM Subject: Histonet Digest, Vol 72, Issue 33 Sent by: histonet-boun...@lists.utsouthwestern.edu 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: a basic question about immunohistochemistry (Thomas Jasper) 2. AW: [Histonet] Biopsy fixation (Nathan Jentsch) 3. RE: SPAM-LOW: RE: [Histonet] a basic question about immunohistochemistry (Patsy Ruegg) 4. AW: [Histonet] Biopsy fixation (Gudrun Lang) 5. Cassette labelling (Anne van Binsbergen) 6. Re: a basic question about immunohistochemistry (Lesley Weston) ---------------------------------------------------------------------- Message: 1 Date: Sat, 28 Nov 2009 10:17:11 -0800 From: "Thomas Jasper" <tjas...@copc.net> Subject: RE: [Histonet] a basic question about immunohistochemistry To: "Salim Yalcin Inan" <syi...@ucalgary.ca> Cc: histonet@lists.utsouthwestern.edu Message-ID: <90354a475b420441b2a0396e5008d4965e3...@copc-sbs.copc.local> Content-Type: text/plain; charset="us-ascii" Dear Salim, As you have been informed, doing immunohistochemistry is possible on this tissue. After all it's possible to do IHC on any tissue whether the conditions you want to test under are ideal or not. Being chastised on this list and calling your work "bad science" is totally out of line and certainly does not help you out. I think some people would do well to reserve judgment, particularly when there's no way they can fully understand what's going on with your project. Having worked in research myself, I completely understand that animals will die, at the most inconvenient times, during a study. First of all you should incorporate the data about the animal dying into your study notes. Secondly, there's no harm in running the IHC on this animal's tissue. You can use the results comparatively with results from some perfused tissue later on. I don't know Salim, some people might call it damage control, or making the best of a less than ideal situation. Again, I don't know exactly what you're working on but it seems there's information worth gathering despite the circumstances. I also understand that it's probably next to impossible to carry out experiments and research alone. Having reliable staff assist you is not unusual either. Good luck to you, Tom Jasper Thomas Jasper HT (ASCP) BAS Histology Supervisor Central Oregon Regional Pathology Services Bend, Oregon 97701 541/693-2677 tjas...@copc.net -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Salim Yalcin Inan Sent: Friday, November 27, 2009 2:44 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] a basic question about immunohistochemistry Dear All, Because I am new in immunohistochemistry, I have a basic question about it. What if your rat dies in the evening or in the weekend, which you are doing a chronic experiment and need to collect brain tissue for immunohistochemistry? And let's say, the staff did not noticed it to inform you on time. Several hours passed since your rat died. There is no way to do perfusion. Is it still possible to do immunohistochemistry? Thank you very much in advance. Best regards, Salim Yalcin Inan, Ph.D. (post-doctoral fellow) Department of Clinical Neurosciences University of Calgary syi...@ucalgary.ca _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 2 Date: Sat, 28 Nov 2009 17:35:31 -0500 From: Nathan Jentsch <njblademas...@gmail.com> Subject: AW: [Histonet] Biopsy fixation To: histonet@lists.utsouthwestern.edu Message-ID: <5a7745af0911281435j339ff9d3ja8d7eff31e931...@mail.gmail.com> Content-Type: text/plain; charset=ISO-8859-1 Gudrun I work in a dermatopathology lab, and we don't see any issues leaving our specimens in 10% NBF for days at a time. We don't work on the weekends, so any specimens that are grossed remain on our processors in NBF from Friday at 8:00 PM until about 5:30 PM on Sunday. I also agree with Kris that 4 hours of fixation is not long enough to ensure proper fixation unless they are shave biopsies. Nate ------------------------------ Message: 3 Date: Sat, 28 Nov 2009 15:55:24 -0700 From: "Patsy Ruegg" <pru...@ihctech.net> Subject: RE: SPAM-LOW: RE: [Histonet] a basic question about immunohistochemistry To: "'Thomas Jasper'" <tjas...@copc.net>, "'Salim Yalcin Inan'" <syi...@ucalgary.ca> Cc: histonet@lists.utsouthwestern.edu Message-ID: <a13fcbaea5394327948047717fe83...@prueggihctechlt> Content-Type: text/plain; charset="us-ascii" Of course you can always do IHC on your rat that died, you just have to note that that rat died and was not profuse fixed so that the review of the results will take that in consideration, hopefully you have another rat that besides dyeing and not getting profuse fixed had all other conditions the mostly the same so you could use the properly treated rat as a standard to compare the dead rat too. Good luck and do keep asking for help at this forum, most of us will offer you our experience without judging your science. Regards, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. Ste.215 Aurora, CO 80045 720-859-4060 fax 720-859-4110 www.ihctech.net www.ihcrg.org -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Thomas Jasper Sent: Saturday, November 28, 2009 11:17 AM To: Salim Yalcin Inan Cc: histonet@lists.utsouthwestern.edu Subject: SPAM-LOW: RE: [Histonet] a basic question about immunohistochemistry Dear Salim, As you have been informed, doing immunohistochemistry is possible on this tissue. After all it's possible to do IHC on any tissue whether the conditions you want to test under are ideal or not. Being chastised on this list and calling your work "bad science" is totally out of line and certainly does not help you out. I think some people would do well to reserve judgment, particularly when there's no way they can fully understand what's going on with your project. Having worked in research myself, I completely understand that animals will die, at the most inconvenient times, during a study. First of all you should incorporate the data about the animal dying into your study notes. Secondly, there's no harm in running the IHC on this animal's tissue. You can use the results comparatively with results from some perfused tissue later on. I don't know Salim, some people might call it damage control, or making the best of a less than ideal situation. Again, I don't know exactly what you're working on but it seems there's information worth gathering despite the circumstances. I also understand that it's probably next to impossible to carry out experiments and research alone. Having reliable staff assist you is not unusual either. Good luck to you, Tom Jasper Thomas Jasper HT (ASCP) BAS Histology Supervisor Central Oregon Regional Pathology Services Bend, Oregon 97701 541/693-2677 tjas...@copc.net -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Salim Yalcin Inan Sent: Friday, November 27, 2009 2:44 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] a basic question about immunohistochemistry Dear All, Because I am new in immunohistochemistry, I have a basic question about it. What if your rat dies in the evening or in the weekend, which you are doing a chronic experiment and need to collect brain tissue for immunohistochemistry? And let's say, the staff did not noticed it to inform you on time. Several hours passed since your rat died. There is no way to do perfusion. Is it still possible to do immunohistochemistry? Thank you very much in advance. Best regards, Salim Yalcin Inan, Ph.D. (post-doctoral fellow) Department of Clinical Neurosciences University of Calgary syi...@ucalgary.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 ------------------------------ Message: 4 Date: Sun, 29 Nov 2009 11:18:52 +0100 From: "Gudrun Lang" <gu.l...@gmx.at> Subject: AW: [Histonet] Biopsy fixation To: "'Nathan Jentsch'" <njblademas...@gmail.com> Cc: histonet@lists.utsouthwestern.edu Message-ID: <355415e1e34e48ee854f2d0a668f8...@dielangs.at> Content-Type: text/plain; charset="iso-8859-1" Nate, I think you have confused the persons. I'm the one, who recommended longer fixation and Kris had made the original question. Regards Gudrun -----Ursprüngliche Nachricht----- Von: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Nathan Jentsch Gesendet: Samstag, 28. November 2009 23:36 An: histonet@lists.utsouthwestern.edu Betreff: AW: [Histonet] Biopsy fixation Gudrun I work in a dermatopathology lab, and we don't see any issues leaving our specimens in 10% NBF for days at a time. We don't work on the weekends, so any specimens that are grossed remain on our processors in NBF from Friday at 8:00 PM until about 5:30 PM on Sunday. I also agree with Kris that 4 hours of fixation is not long enough to ensure proper fixation unless they are shave biopsies. Nate _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Sun, 29 Nov 2009 16:24:20 +0400 From: Anne van Binsbergen <anni...@gmail.com> Subject: [Histonet] Cassette labelling To: "histonet@lists.utsouthwestern.edu" <Histonet@lists.utsouthwestern.edu>, histonet-requ...@lists.utsouthwestern.edu Message-ID: <f8332fbe0911290424w2c0f8a9dx1b0214168cc6a...@mail.gmail.com> Content-Type: text/plain; charset=ISO-8859-1 Hi Histonetters Who out there has a Sakura iDent? I have just taken delivery of a brand new machine and I am having serious issues with the quality of the print on the cassette face - the ink rubs off with xylene!!! Yes, I leave the print to dry before I handle the cassettes No, I dont use Tissue-Tek xylene substitute - my xylene is made by Fisher I would love to know if anyone else has had similar issues Yes, Rene, this is the diehard number 1 Sakura fan having issues with a Sakura product!!! hopefully yours (pun intended) AbuDhabiAnnie -- Anne van Binsbergen (Hope) Abu Dhabi UAE ------------------------------ Message: 6 Date: Sun, 29 Nov 2009 06:22:57 -0800 From: Lesley Weston <les...@shaw.ca> Subject: Re: [Histonet] a basic question about immunohistochemistry To: histonet@lists.utsouthwestern.edu Message-ID: <bf6c7b78-0305-48e3-a6e2-584be0e2c...@shaw.ca> Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed Just about all biology research institutions have an animal unit with qualified technicians to look after the animals, rather than someone qualified in other areas; however the technicians are subject to human failings such as going home at night. I've never heard of a one- person research team, and I don't think it would be all that effective. The tissue from the animal that died at the wrong time is not comparable to perfused tissue, but as Tom says, it might still be worth processing. If nothing else, it will show how much difference perfusion makes. Lesley Weston. On 28-Nov-09, at 10:17 AM, Thomas Jasper wrote: > Dear Salim, > > As you have been informed, doing immunohistochemistry is possible on > this tissue. After all it's possible to do IHC on any tissue whether > the conditions you want to test under are ideal or not. > > Being chastised on this list and calling your work "bad science" is > totally out of line and certainly does not help you out. I think some > people would do well to reserve judgment, particularly when there's no > way they can fully understand what's going on with your project. > Having > worked in research myself, I completely understand that animals will > die, at the most inconvenient times, during a study. > > First of all you should incorporate the data about the animal dying > into > your study notes. Secondly, there's no harm in running the IHC on > this > animal's tissue. You can use the results comparatively with results > from some perfused tissue later on. > > I don't know Salim, some people might call it damage control, or > making > the best of a less than ideal situation. Again, I don't know exactly > what you're working on but it seems there's information worth > gathering > despite the circumstances. I also understand that it's probably > next to > impossible to carry out experiments and research alone. Having > reliable > staff assist you is not unusual either. > > Good luck to you, > > Tom Jasper > > > Thomas Jasper HT (ASCP) BAS > Histology Supervisor > Central Oregon Regional Pathology Services > Bend, Oregon 97701 > 541/693-2677 > tjas...@copc.net > > -----Original Message----- > From: histonet-boun...@lists.utsouthwestern.edu > [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Salim > Yalcin Inan > Sent: Friday, November 27, 2009 2:44 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] a basic question about immunohistochemistry > > Dear All, > > Because I am new in immunohistochemistry, I have a basic question > about > it. > What if your rat dies in the evening or in the weekend, which you are > doing a chronic experiment and need to collect brain tissue for > immunohistochemistry? And let's say, the staff did not noticed it to > inform you on time. Several hours passed since your rat died. There is > no way to do perfusion. Is it still possible to do > immunohistochemistry? > Thank you very much in advance. > > Best regards, > > Salim Yalcin Inan, Ph.D. > (post-doctoral fellow) > Department of Clinical Neurosciences > University of Calgary > syi...@ucalgary.ca > > ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 72, Issue 33 **************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet