[Histonet] osteoblast count
HI I have a problem with counting osteoblast in IHC staining slides is there a way to differentiate osteoblast from other bone cell types. thank you very much Hatem ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Processing Whole prostate
Hello to all in histoland. For those who are doing whole mounts for prostate, be willing to share the process with me. My lab is looking into to doing this. I would need to know the euipment used and processing times. Your help in this matter would be greatly appreciated. Allison Scott HT(ASCP) Histology Supervisor LBJ Hospital Houston, Texas 77026 7135665287 CONFIDENTIALITY NOTICE: If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail and any attachments from your computer system. To the extent the information in this e-mail and any attachments contain protected health information as defined by the Health Insurance Portability and Accountability Act of 1996 (HIPAA), PL 104-191; 45 CFR Parts 160 and 164; or Chapter 181, Texas Health and Safety Code, it is confidential and/or privileged. This e-mail may also be confidential and/or privileged under Texas law. The e-mail is for the use of only the individual or entity named above. If you are not the intended recipient, or any authorized representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail and its attachments is strictly prohibited. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Nuclear DAPI staining in frozen sections of bones
I am doing nuclear DAPI staining in frozen sections of fresh, non-decalcified mouse femurs. The fresh femurs were snap frozen in OCT block by 2-methylbutane (isopentane)/dry ice. Cryosection of the bones were done by using the Instrumedics CryoJane tape transfer system. After transferring sections to slides, the slides were kept in -80oC until staining. But I have problem with the quality of nuclear DAPI staining, although HE staining is pretty good. The nuclear DAPI staining (Vectorshield with DAPI or 1 uM DAPI, 10 min in PBS) is fussy and unclear. It is hard to identify each individual cell by nuclear staining. If anyone else has experience on DAPI staining in frozen bone sections, could you share any suggestion to improve the DAPI staining? Any help would be greatly appreciated. Shin-Young Park ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] osteoblast count
I am not sure about after staining with IHC, but if you stain with a toluidine blue or MacNeal's tetrachrome you can easily identify osteoblasts. Jack Date: Fri, 16 Oct 2009 07:56:40 -0700 From: dr.hatemsa...@yahoo.com To: histonet@lists.utsouthwestern.edu Subject: [Histonet] osteoblast count HI I have a problem with counting osteoblast in IHC staining slides is there a way to differentiate osteoblast from other bone cell types. thank you very much Hatem ___ 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] Perfect perfusion
Hi Everyone!! Is there a way to get perfect perfusion for each mouse every time? I am getting variable perfusion quality for each mouse. I am perfusing CO2 euthanized 3-4 weeks old mice with PBS via peristaltic pump followed by fixative (RNAlater) via syringe merging into the same tubing for PBS. I am nicking the upper right atrium of heart to collect the gushed out blood and then perfusing through ventricle using a 21G butterfly needle. I am collecting brain and spinal cord. Thanks much, Dzung NIAID ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Rabbit-on-Rodent HRP polymer
Jennifer, You will not need to use the avidin/biotin block in your protocol. The polymer technology does not use any sort of biotinylation or avidin system for staining. That should help knock some time off your protocol. You will need to make sure you use a peroxidase block (3% hydrogen peroxide) in your staining protocol to block endogenous peroxidase activity (namely red blood cells/sera). Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Rabbit-onRodent HRP Polymer
Hi Jen, The rabbit-on-rodent HRP ploymer is ready-to-use. In my protocol I depar and hydrate to wash buffer then block for endogenous peroxidase for 10 minutes using Dako's DEEB followed by wash buffer rinses then block for 15 minutes in Rodent Block M (Biocare). No need to block for biotin since no streptavidin is used in the detection. I suggest that you first try the block you have. Apply rabbit primary antibody for 30 minutes, rinse in wash buffer, then apply the rodent-on-rabbit HRP polymer for 15 to 30 minutes. Rinse in wash buffer again then apply your DAB chromagen for 5 minutes. Rinse well in DI water then stain with hematoxylin. I run all of my IHC on a Lieca Bond Max using the Biocare reagents and they work great. This protocol intended for general guidance; the exact conditions will need to be worked out in your lab. Hope this helps. Dave Jen wrote: Message: 11 Date: Thu, 15 Oct 2009 17:36:41 -0700 From: Jennifer Campbell jcampb...@vdxpathology.com Subject: [Histonet] Rabbit-on-Rodent HRP polymer To: histonet@lists.utsouthwestern.edu Message-ID: 5658cbdb9eae6545abe50d2563d81bf8181...@vdxserver01.vdxpathology.local Content-Type: text/plain;charset=iso-8859-1 Hi All, I recently purchased a rabbit-on-rodent HRP polymer from Biocare. I will be using this on mouse tissue, using a rabbit polyclonal CD3 as my primary. I just wanted to clarify a couple things about the protocol before I try it out tomorrow and was wondering if anyone could offer some assistance. First of all, I should aske if anyone has a protocol for this? What I really would like to make sure is that I will not have to use a label after the rabbit-on-rodent secondary ab. I believe that I will just have to apply my blocking agent, avidin/biotin blocks, primary antibody, secondary anitbody, followed by chromogen, right? My blocking agent is made up of casein and some other proprietary agents in a phosphate buffer (it is non-serum) so I was told that should work fine. Does this sound correct? Thanks in advance! I haven't been able to get through to tech support all day, so hopefully you will have some answers for me! Jen ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Oil Red O protocol
I¹m looking for a protocol for Oil Red O that does not use propylene glycol to make the Oil Red O. Does anyone have a protocol that I could use? Aprill Watanabe, B.S. Research Associate Integrated Cancer Genomics Division Tissue Microarray Center (TMA) Translational Genomics Research Institute (TGen) main: 602-343-8822 Fax: 602-343-8840 awatan...@tgen.org www.tgen.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Saturday Tech Needed- Orange County California
Hello, Private lab seeking a histoch to work on Saturdays. Will be rotating with the other Saturday techs. Shift starts at 5 or 6 am. If interested, please send me an email. Thank you, Paula Lucas Lab Manager Bio-Path Medical Group Fountain Valley, CA 92708 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Need maintenance/service on Microm HM340E-2 microtome in Waltham, MA please
Hello Histonet, I am looking for a company other than Thermo Fisher to do annual maintenance and repairs on our Microm HM340E-2 microtome. Please email or call me as soon as possible. Emailing your price list would be very helpful. Here are the details: It is 2 years old but virtually brand new (hardly used until recently). It's been working fine, then I moved it from one room to the next yesterday and today I am getting a strange venetian blind pattern (first section ok, second section ok, 3rd section bottom third venetian blind, 4th section bottom half venetian blind, 5th and 6th sections all venetian blind and then it's fine again and it repeats the pattern all over). I'm using a paraffin block with no tissue in it to trouble shoot. Section thickness: 4 microns Blades: DuraEdge high profile, same package that was working fine, new blade (tried 4 different ones) Angle: 8 degrees (unchanged)-tried 10 and it made no difference Cold block: yes -I checked if everything was tight. -I tried the blade holder tightener at 3 different spots. When I loosened it I got thick and thin sections. Nothing feels loose! I think whatever is wrong is inside (a spring, or the mechanism that keeps section thickness at 4 microns-at 5 microns the pattern seems to go away??) I look forward to your responses. Thank you and Happy Friday. Sharron Ladd Senior Histotechnologist On-Q-ity 610 Lincoln Street, 3rd Floor Waltham, MA 02451 Phone: (781) 895-8110 Fax: (781) 890-4636 sharron.l...@on-q-ity.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
