THE CENTRAL MICROSCOPY FACILITY
Antifade Mixtures for Fluorescence
Microscopy are media for mounting preserved samples that will be
viewed by fluorescence microscopy. These mixtures serve several
- provide a mounting medium that is compatible with preserved
tissues and cells,
- glycerol-based mounting medium has an index of refraction
that is appropriate for some objectives, and much better than aqueous
- enhance long-term storage without loss of specimen quality
- slow the photobleaching rate when the specimen is observed.
There are do-it-yourself simple formulations that are inexpensive and
easy to prepare, and there are a number of proprietary products that
may have specific advantages for their stated purpose. For example,
some media give a lower initial fluorescence intensity, but the
subsequent rate of photobleaching is much lower, giving a more stable
fluorescence for measurements.
These mixtures are prepared by simply mixing the ingredients until
dissolved (several hours to overnight) and filtering or centrifuging.
They are stored dark at room temperature (or colder, to -20C), and
appear to be stable for several years. A search of the Microscopy
America Listserver Archives will find a number of
contributions on this subject. From that source, Peter Sargent
(comparing DABCO, phenylene diamine, n-propyl gallate,
Vectashield, and Slow Fade) determined that 4% n-propyl gallate in 90%
glycerol and 10% PBS was the best he tested; slides are reputed to keep
for weeks to years in dark at -20C.
- 4% n-propyl gallate (Sigma) in 90%glycerol, 10% phosphate
buffered saline (PBS)
- 1% DABCO (Sigma) in 90% glycerol, 10% phosphate buffered saline
Probes >> http://www.probes.com
ProLong(r) Antifade Kit - claimed to be best by Molecular Probes
Slow Fade(r) - slides keep 2-3 months in dark @ 4 C.
Laboratories >> http://www.vectorlabs.com/
Vectashield, Vectashield Hard-set
The Central Microscopy
Facility >> http://www.bio.umass.edu/microscopy
The University of Massachusetts at Amherst, MA, USA