I am trying to optimize mounting media for IF stains. Vectashield and Prolong are both very good, but very expensive. That is why I am thinking of making a "home-made" custom blend to reduce the costs.
As my primary source I read-up about various options on this website:
http://www.ihcworld.com/_protocols/histology/aqueous_mounting_medium.htm
In that list Buffered glycerol with anti-fade, seems the best option so I am going with that.
It lists two anti-fades as a viable option:
- p-phenylenediamine (PPD)
- n-propyl gallate (NPG)
A third option might be Ascorbic acid, which as an antioxidant might have similar effects.
Does anyone have any experience with these?
Is there any use in mixing different anti-fades together to get a better result, or is it best to stick just to one?
PPD is often mentioned as the best option, how does it work with DAPI, if DAPI is added directly to the mounting medium?
Without being aware of what your cost line is, I'd recommend fluoromount G (http://www.emsdiasum.com/microscopy/products/histology/mounting_media.aspx#17984). It works very well in my hands and is not very expensive.
I have no experience making my own, but I imagine it's more economical for your time to have a pre made solution instead of making your own.
I changed to Fluka #10981 PVA with DABCO. Comes in a 100mL bottle for approx 120 euros; cheap compared with those small flasks of similar products. Works fine for us working with cells and tissues using Alexa, Cy, and NL-dyes, with DAPI as well. Might be not the same quality as "Prolong" but well enough for most applications.
If the medium becomes too thick, transfer an aliquot to an Eppendorf tube, heat it it for some minutes @ 50-60 °C and it will liquefied again.
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