[Histonet] HCN4 staining
Hello, I'm hoping somebody out there can give me some help with HCN4 staining, I was asked to stain HCN4 on mouse heart and it needs to stain the SA node. It is very hard to find that area since the node is very small. I have tried serial sections but not sure I hit the area or if I'm having problems with the stain. The antibody is Rat monoclonal Secondary Alexa Flour 488 goat anti-rat Counter stain is Hoechst I tried frozen and paraffin sections with no luck Any suggestions will help greatly Thanks, Sharon ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Rat Skin
Can anybody share their protocol on processing rat skin I seem to be having a problem with it being under processed. I fix the tissue for 11hrs in zinc formalin and then run a 12 hour processing schedule the skin size is 2.5cmx1cm. Thanks, Sharon ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Starting up new lab
I'm looking at starting up a new histology lab and need everything, I have a tight budget. Can anybody give me recommendations on where to buy refurbished equipment. Thanks Sharon ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] refurbished equipment
Hello everyone, Vendors and sales reps please don't call my office and leave messages it is a shared phone and this is an opportunity outside my current position please leave me an email and I will get back to you. Many Thanks, Sharon ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Rat skin Staining
Hello, I'm looking to see if anybody has HE transverse sections of rat skin that you are willing to send me a copy with the skin sections labeled so I can see what I'm looking at on my sections. Thanks, Sharon-WPI ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] mice legs
Good Morning Histo World, I would like to know if anyone is working with mice legs, I have a PI that I work with that wants to process the whole leg, the problem is I need to decal it first and is wondering if the decal will break down the tissue, I think it would he doesn't think so. And if anyone has do this would it be possible to share your protocol with me from decal to processing. Thanks, Sharon- WPI ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] DNA Extraction
Hello Histo land, Does anyone know how to extract DNA from formalin fixed paraffin embedded tissue? Thanks, Sharon WPI ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Decal Solution
Hi Histo world, I have a crazy question, I work in research facility and one of the investigators needs decal done on mice legs, they never check to see if we had decal solution before they began the procedure, of course we don't. I tried different companies to order some but they can't overnight the solution because it is a hazard. Does anyone have a protocol that they use in making a decal solution? The old histology books do but I was looking for maybe an updated solution. Please send my your suggestions they want this done ASAP. Thanks, Sharon Shaw WPI ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Anti-NuMA (Ab-2) Mouse mAb (107-7) Calbiochem cat no. NA09L
Hello, I have recently began using the Anti-NuMA primary antibody to distinguish the human mesechymal stem cells that I am delivering to the rat heart. Anti-NuMA recognizes human nuclear mitotic apparatus protein. I have tried two secondary antibodies: Alexa Fluor 488 and FITC (Jackson immunoresearch code 715-095-150) and have positively stained human mesenchymal stem cells put on glass coverslips with both secondary antibodies. However, when I look at the sections of my rat heart that I have delivered human mesenchymal stem cells to, I get a lot of autofluorescence and no positive signal (counterstained with Hoechst and the hMSCs are Quantum Dot loaded). The sections were fixed in 4% paraformaldehyde right after the animal was euthanized and heart removed. The heart was then frozen in OCT and sectioned. I have tried changing the concentration of both the secondary's and primary's already (within the manufacturer's specs). My question is: what secondary antibodies do you recommend? Does anyone have experience with this primary? Thanks, Sharon WPI ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] EDC Fixation
Hi Everyone, I'm looking to see if anyone has used EDC fixation with formalin, if so do you EDC fix first followed by formalin or formalin fix followed by EDC. I would love to have some feedback on this. Regards, Sharon ___ Sharon Shaw Worcester Polytechnic Institute Laboratory and Histology Manager Department of Biomedical Engineering 100 Institute Road Worcester,MA 01609 508-831-4125 shs...@wpi.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
