Hi,
I want to start using an IHC protocol based on PBS and triton.
I have a DPX mounting medium, would it be OK to use? or should I have an aquatic-based one? I don't want bubbles and of course, I don't want to harm the staining.
thanks!
Shalom Nitzan,
What staining technique are you going to use?
If you are looking at immunofluorescence IHC, then DPX is not suitable. Immunofluorescence samples are very delicate compared to basic staining techniques. you need to use a medium that will prevent the IF signal from fading. I suggest looking at Vectrolab's catalogue of IF mounting media (vectorlabs.com).
If you are using light microcopy then immunofluorescence then DPX should be fine but you need to dehydrate the tissues (take down to water) before mounting, which can affect tissue morphology.
In this case you could use Limonene mounting medium which is aqueous and works well; you can find it in abcam's catalogue (abcam.com).
Don't forget to seal your slides with clear nail varnish!
Good luck and have fun with IHC.
Hello,
if you use aqueous light microscopy staings or immunofluorescence then you are not allowed to use DPX!
If you use a final viusalization with Diaminobenzidine (DAB), then DPX (after dehydration with graded alcohols (from 50-100%) and finally 3 x 10 min in Xylene) can be used and is a very good mounting medium.
Kind regards
Stefan
Since you wrote IHC protocol, considering the HRP DAB method, DPX is very good mounting media, of course after dehydrating and clearing the slides.. Regards
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