Selection of the appropriate product:

There is no universal antifadent due to the fact that biological material is so varied in nature that the fluorescence fading of fluorochromic conjugates is a complex process.

AF1, AF2 and AF3 are general antifadents. AF87 is a specialist immersion-oil-based antifadent, which can be used with specimens after they have been dehydrated.  The CFPVOH/AF100 mixture is a gel, which may be used in this form, provided the edges of the cover slip are sealed with nail varnish.  If the edges of the cover slip are not covered, the solution dries to give a clear film immobilising your specimen.  Products, which may be used for the same purpose, include the CFPVOH plus antifadent and the Permafix PF plus either AF200 or AF300.  The Permafix PF plus AF300 cures to give a good hard clear film, firmly attaching the cover slip to the specimen and slide.

AF1 should be used if you normally examine specimens in glycerol solution. If you normally use saline solutions with your specimens AF3 is appropriate. AF2 is simply a glycerol solution of the antifadent.

Obtaining the correct viscosity

If the viscosity of the AF1 mountant solution is too high for your application you can always reduce the viscosity by adding some AF3 solution. Both AF1 and AF3 contain the same antifadent at comparable concentrations and hence mixtures of the two solutions may be safely used. The chart below gives some indication as to how the viscosity varies with the percentage of AF1 in the mixture.

If you think this trick may be of value, why not order 1X25ml AF1 together with 1 X 25ml AF3 and obtain a discount? Details on Orders pages.

Increasing the amount of AF2 solution increases the viscosity:

If samples AF1, AF2 or AF3 do not meet your needs, please contact us. We have experience with many other systems and may be able to supply a product which meets your requirements.

AF87. The antifadent is contained in a nonfluorescent immersion oil. It is designed for those requiring work at a high magnification. It is particularly useful for preventing the fading of such dyes as DAPI which have the unfortunate tendency to disperse in glycerol based mountant solutions.

CFPVOH is an aqueous solution of poly(vinyl alcohol) for use as a solid mountant. Apply a few drops of the solution to your specimen and allow to dry to give a clear film.

AF100 is a solution of antifadents to be used with CFPVOH when fading of fluorescence in the poly(vinyl alcohol) is a problem. To prepare a CFPVOH solution containing AF100, add 0.1ml. of AF100 to 0.9ml. of CFPVOH. If more or less of the solid mountant solution is required, mix the reagents in the appropriate ratio. Once these solutions have been made up, they should be used within a period of days rather than months since the solutions deteriorate on storage.

Some important facts about our solutions AF1, AF2 and AF3 These products have a long shelf life. AF1 solutions have travelled around in the back of a car and still work well after 2 years. However you should check that any deterioration that does occur on storage will not adversely affect your results. The solutions can be stored at room temperature i.e. there is no need for refrigeration and if you leave the cap off the bottles by mistake, the solution doesn't lose its efficacy over a short period of time. The performance of the products is dependent on the nature of the biological material being studied and consequently the efficiency of preventing photofading is substrate dependent. The kinetics of signal loss are substrate dependent. When a large group of substrates is considered the Citifluor Solutions have a very high success rate. The LD50's for these three products is >2000 mgm/kgm (in rat).  Provided good housekeeping is exercised, none of these products should pose toxicological problems AF1, AF2 AF3, AF4, PVP, MWL are all supplied in plastic bottles.  Note: these products can be supplied in brown glass bottles but this will lead to higher postage costs. AF100, AF200, AF300, CFPVOH, CFPVOH plus antifadent, the Permafix, CFM and UVM solutions are all supplied in glass bottles. Larger pack sizes can be supplied upon request – ask us for a quote for the amount of material you require. Prices can be found on the Orders pages.

Green Snow Algae mounted in antifadent solution AF2 Courtesy Dr. D. Wynn-Williams, British Antarctic Survey

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