[Histonet] pancreas pH or proteases inhibit LacZ enzyme activity?

Hello, Histonet experts, I have a puzzling bit of data that I am trying to resolve. Our group and another group have performed LacZ stain on mouse pancreas to identify cells expressing a LacZ reporter (the other group has published this expression pattern). I am having trouble with the primary

[Histonet] immunofluorescence mounting medium?

Hello, Esteemed Histonetters, I am trying to get nice publication-quality images of my immunofluorescent tissue sections. I am currently using Prolong Gold, and after I let the stuff cure for several days, my 100X oil-immersion images are still smeary, even after taking into account the

[Histonet] RNAlater regret

Hello, Esteemed Histonetters! I have a collaborator who asked me to drop his (mouse) bones into RNAlater for RNA isolation, later. These animals were double labeled for histomorphometry as well. He now wants to know if he can back them out of RNAlater and use them for any histology-related

[Histonet] Frost papers on bone labeling

Hello, Esteemed Histonetters, I am wondering if anyone happens to have a 21st century version of a 20th century paper? In Search Of PDF files for any /all of the following: Frost HM. Presence of microscopic cracks in vivo in bone. H. Ford Hosp. Med. Bull. 1960;8:25–35. Frost HM, Villanueva

Re: [Histonet] Manual embedding

Hi, All, I just saw this question and the responses, thought I would add my own solution to the mix. I do manual embedding, I use a hyb oven for my infiltrating/embedding station (like for hybridizing Southern or Northern blots- who does that anymore?). I have taken out the rotating wheel for

[Histonet] Plastic processing for undecalcified mouse bones

Hello, All, I am looking into starting a plastic embedding/sectioning endeavor, and am trying to figure out my startup reagents and protocols. I am seeing a lot of protocols for the Technovit system, which uses a slide press to adhere the sections to the slides, and would prefer to avoid

Re: [Histonet] Question from one of our researchers

Hi, Teri, How about this? It also gives a brief blurb in the description about commonly used alternatives, if this isn't your thing. I've never tried it, so I can't vouch for it myself, but Invitrogen does make some pretty darn good stuff... http://probes.invitrogen.com/media/pis/mp10045.pdf

[Histonet] Blade sharpening service

Hello, esteemed colleagues, I am in need of a service to have my 16cm tungsten carbide Profile D (a.k.a. Sweeney Todd) blades resharpened. Can anyone recommend a service that is relatively accessible to the West coast? I looked in the archives and couldn't find any recent posts, and my Googling

[Histonet] strontium vs. calcium in bone

Hi, Everyone, Happy Friday! I was wondering if anyone knows of a histological method to distinguish strontium in bone vs. calcium? My understanding is that mineral stains (von Kossa or Alizarin Red S) do not distinguish, but was wondering if anyone has a procedure I may not have done or seen?

RE: [Histonet] Coverslipping Problem - Please Help!

Hi, Brian, It is likely to be a pH issue. If you use BM Purple as your detection chromogen (which is what we use in our lab for whole mounts on X. laevis and X.tropcialis, and mouse embryos for WMISH), then you must keep the embryos at acid pH from BM Purple step onwards, or you will

RE: [Histonet] Water collecting at bottom of sections

I use FisherBrand SuperFrost Plus charged slides, the only time I have this problem is when my waterbath is too hot, or if for some reason I use warmed slides to retrieve my sections from the bath (like if you lay the unused slides on the edge of the waterbath, or if you use the warm droplet

[Histonet] RE: X-gal staining troubleshooting

Hi, Teri, I have seen it in some of my stains, but they were always in embryos of mice from a particular strain (that we did not generate and have incomplete construct information for). I always attributed it to a different LacZ allele, as I have stained litters of a different strain