Re: [Histonet] linear stainer hematoxylin staining issues
How do I unsubscribe from Histonet?? Thank you, Arjun On Wed, Mar 27, 2013 at 1:46 PM, Kienitz, Kari kkien...@orclinic.comwrote: If your acetic acid solution is made in-houseI would suggest remaking it, just in case it was accidently made stronger. Kari Kienitz HT, (ASCP) Histology Laboratory Portland Gastroenterology The Oregon Clinic NE 99th Ave Portland, OR 97220 503.935.8311 kkien...@orclinic.com CONFIDENTIALITY WARNING: This e-mail and any attachments are for the exclusive and confidential use of the intended recipient. If you are not the intended recipient, please do not read, distribute or take action in reliance upon this missive. If you have received this in error, please notify the sender immediately by reply e-mail and delete this message and its attachments from your computer system. Thank you From: histonet-boun...@lists.utsouthwestern.edu [ histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sophia Lin [ lins0...@gmail.com] Sent: Wednesday, March 27, 2013 9:26 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] linear stainer hematoxylin staining issues We have the Hacker linear stainer, and for the past two weeks we had clean, crisp HE staining. However, last Friday, something happened and our HE stains no longer look crisp, but instead have a hazy, unreadable look. Nothing different has been done, and we have troubleshot for every possible scenario. After a clean set up, we noticed that the hematoxylin is not staining very well. We have it set for 2 minutes with agitation for a uniform staining. Hematoxylin is Harris Special (mercury free) from MCC and the eosin is with phloxine b from AMT. After the hematoxylin (2 buckets for total of 4.5 minutes), we use 1.5% acetic acid. It seems that the hematoxylin is not staining well. Any tips? We may need to consider increasing a bucket for hematoxylin? Thanks, Sophia ___ 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] How to fix frozen section?
i use 1:1 Acetone : alcohol. On Nov 6, 2012, at 2:41 PM, Amy Lee wrote: Hello, I will receive some frozen tissue sections that are not fixed. I rarely do IHC on frozen sections. I searched some articles about fixation. Some use 50/50 of acetone/alcohol and some use only alcohol. Could you please recommend a fixation method for me? Thanks, Amy ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Best, Arjun ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Fwd: MIXED fiber type identification by Immunohistochemistry
Dear all, Its been a while i asked for a doubt to all histology experts around US, but i have not been able to get much reply (only 1 :(). I thought my doubt/e mail might have been missed by some, so sending it again. I would really appreciate if anybody could guide me in my problem. Hope to hear people's advices this time. Thanks Best, Arjun Begin forwarded message: From: arjun ruhella ruhella.ar...@gmail.com Date: October 2, 2012 8:34:59 AM EDT To: histonet@lists.utsouthwestern.edu Subject: MIXED fiber type identification by Immunohistochemistry Greetings to all, In immunohistochemistry, I am encountering confusion in designating muscle fibers (based on staining color intensity) to a particular fiber type. This problem is particularly bothersome when i encounter the fibers stained with 2 different colors, one color has good intensity while other seems to be of light intensity. I don't have any criterion whether to consider this light staining as actual staining, which would classify the fibers as MIXED fibers or should i consider this light staining as background/non specific staining in which case the fibers will be classified as PURE fibers. Does anybody can suggest any criterion/tips they have been using to identify muscle fiber type as correctly as possible? I can understand that identifying MIXED fibers is subjective in immunohistochemistry, but it would be of immense help if i could get to know of any objective suggestions. Animal= spinal cord injured Rat Muscle = Soleus Section Thickness = 10µm I will really appreciate the help! Thanks, hope to hear from ppl soon! BEST, AR Best, Arjun ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Mixed fiber type identification
Thanks Michael! I really appreciate your suggestions. That being said, i really cannot go for mATPase or PAS stain, I need to do IHC. I hope to hear from other too on this issue. Thanks AR On Oct 4, 2012, at 1:20 PM, mtitf...@aol.com wrote: Arjun Ruhella asked about fiber typing of animal soleus muscle: No one answered him, at least in public. I guess all the hotshots are in Vancouver! Years ago I was involved in fiber typing and measuring of human muscle biopsy fibers. In those human muscle biopsies, all the bundles were mixed, about 60:40 if I remember correctly. The proportion and size of the fibers changed depending on the disease process. We used the ATP reaction on frozen sections to visualise the type 1 and type II, (and IIA, IIB, etc) fibers. Then, in those pre-computer days, we selected a typical muscle bundle and started counting the cross sectional diameter of those type I and type II fibers, using eypepiece graticule in the microscope. A histogram was prepared. I cannot imagine using IHC to demonstrate the type I and Type II fibers is a whole lot different, or that animals have muscle fibers that are not mixed. You can see some differentiation on a good PAS stain, but I do not know how well it compares. The procedure we used was in Muscle Biopsy: A modern approach by Dubrowitz and Brooke, Saunders. 1973 ( I told you it was years ago!) Hope this helps Michael Titford Pathology - USA Mobile AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Best, Arjun ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] MIXED fiber type identification by Immunohistochemistry
Greetings to all, In immunohistochemistry, I am encountering confusion in designating muscle fibers (based on staining color intensity) to a particular fiber type. This problem is particularly bothersome when i encounter the fibers stained with 2 different colors, one color has good intensity while other seems to be of light intensity. I don't have any criterion whether to consider this light staining as actual staining, which would classify the fibers as MIXED fibers or should i consider this light staining as background/non specific staining in which case the fibers will be classified as PURE fibers. Does anybody can suggest any criterion/tips they have been using to identify muscle fiber type as correctly as possible? I can understand that identifying MIXED fibers is subjective in immunohistochemistry, but it would be of immense help if i could get to know of any objective suggestions. Animal= spinal cord injured Rat Muscle = Soleus Section Thickness = 10µm I will really appreciate the help! Thanks, hope to hear from ppl soon! BEST, AR ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] How can i post a question in histonet?
Please let me know! Thank you Best, AR ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] How can i post a question in histonet?
Oh, i see. I did not know this is how it works. Thank you. Hope to gain knowledge from this site. Best, AR On Sep 20, 2012, at 1:50 PM, Connolly, Brett M wrote: You just did. Brett -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of arjun ruhella Sent: Thursday, September 20, 2012 1:49 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] How can i post a question in histonet? Please let me know! Thank you Best, AR ___ 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
[Histonet] Is specimen too cold for cryosectioning?
Dear all, I have heard a little trick in cryosectioning where, if the specimen is too cold, one put thumb on the specimen and then cut immediately. I have a question related to this. Q1 = My cryostat temperature cannot be increased above -20C (a problem needs to be fixed). I usually tend to cut my specimen at ~-18C. As i was bound to cut the specimen at -20C, i was experiencing chattering or splintering in the sections. I though this was because the specimen was very cold. When i rub my thumb on the specimen for a while, i was able to get 2-4 decent sections, but the chattering started again. Is this a CONCRETE evidence that the chattering is due to specimen being cold? Q2 = Does sectioning difficulty can happen with as small difference as -2C (-18C vs -20C)? I heard ppl saying that -2C will not make any difference in sectioning. Hope to hear from someone soon! Sincere thank you! AR ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
