Various Mounting Media and Anti-fade Reagents:
(Not a complete listing. This is a collection of suggestions from many users. Please contact me with your suggestions, corrections, and additions.)
Mounting Media:
The sample's surroundings will affect the image to a great degree. For optimal quality, you would ideally like your sample to be immersed in a material with the same index of refraction as our immersion oil (1.518). If the material is able to solidify, you will avoid problems with movement of the sample (or the coverslip) relative to the slide. And for unstable fluorophores, which tend to photobleach quickly, many of the commercial mounting media contain anti-fading reagents in them.
Note that your sample's characteristics will determine whether you want an aqueous medium or an organic one.
- PermaFluor from Lipshaw:
(aqueous, sets hard)
- FluorSave:
(aqueous)
- ProLong B from Molecular Probes:
(sets hard)
- Fluoroguard from Biorad
- VectaShield from Vector Labs
- PolyMount or Aqua-PolyMount from Polyscience
- MOWIOL from Calbiochem
- DPX
- 50% glycerol, 50% PBS
(homemade, buffered to correct pH for your fluorophore)
- methyl salicylate
(organic, dissolves plastic and nail polish, but has excellent optical properties)
Anti-fade additives:
Many people prefer to add specific anti-fade reagents, such as the following, to their mounting media (often made from scratch). Although the mechanism of the photobleaching has not been completely elucidated, it has been found that you can reduce the effect by using free-radical scavengers, minimizing the amount of oxygen in the sample, etc.
- 1,4-diazobicyclo-(2,2,2)octane (DABCO)
- n-propyl gallate
- p-pheylenediamine dihydrochloride
- ascorbic acid
Keep in mind that every fluorophore has an optimal pH range, and buffer your sample accordingly.
JHS
03/25/99 |
