[Histonet] Jones problems

[Amos Brooks]

Hi,
     I think your silver precipitation issue could be cured by the
thiosemicarbizide step too. If the reaction is sped up by this, you won't
need to have the slides in long enough to develop the precipitate.
    That said, since it is a silver stain, you still need the glassware to
be acid cleaned, but what you've been doing is probably just fine. I'm
pretty sure this will solve your problem. Keep us posted though.

Cheers,
Amos

On Thu, Oct 7, 2021, 12:57 PM Hood, Jordan <jordhood at med.umich.edu> wrote:

> Hi Amos,
>
>
>
> It definitely helps, thank you! I brought this to my pathologist and she
> said that it makes complete sense. I plan on ordering some TSC in solid
> form.
>
>
>
> Is there a chance that I didn’t do the acid-washing of the glassware
> correctly, leading to contamination? Or could all of the precipitate
> floating around on the warm silver solution and on the slides be due to the
> instability of the silver solution?
>
>
>
> Thank you so much!
>
> Jordan
>
>
>
> *From:* Amos Brooks <amosbrooks at gmail.com>
> *Sent:* Saturday, September 25, 2021 8:03 AM
> *To:* Hood, Jordan <jordhood at med.umich.edu>;
> histonet at lists.utsouthwestern.edu
> *Subject:* Jones problems
>
>
>
> *External Email - Use Caution *
>
> Hi,
>
>      When I first did this stain, I had really light staining of the GBM.
> StainsFile has a note in the procedure that describes thiosemicarbizide.
> Being a complete nerd, I wanted to see another source as well. Here is a
> link to an article where it is used...
>
> https://pubmed.ncbi.nlm.nih.gov/2482996/
>
>      I also make these chemicals freshly each time. I accomplish this by
> pre-weighing the solids sufficient for a 50ml volume and storing it in an
> eppendorf tube. When you need it, pop the tube into the requirement volume
> of water and you're golden.
>
>      I do this because in addition to having good freshly made solutions,
> I found the thiosemicarbizide goes bad rather quickly after use. TSC is the
> lynch pin for the procedure. The silver on it's own does not react very
> strongly with the aldehydes that are reduced by the periodic acid without
> the TSC to facilitate the precipitation.
>
>
>
> I hope this helps,
>
> Amos Brooks
>
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