Fluorescent biosensor molecules, fluorescent biosensors and systems,
as well as methods of making and using these biosensor molecules and systems are
described. These biosensor molecules address the problem of obtaining fluorescence
emission at wavelengths greater than about 500 nm. Biosensor molecules generally
include an (1) an acridine-based fluorophore, (2) a linker moiety and (3) a boronate
substrate recognition/binding moiety, which binds polyhydroxylate analytes, such
as glucose. These biosensor molecules further include a "switch" element that is
drawn from the electronic interactions among these submolecular components. This
fluorescent switch is generally "off" in the absence of bound polyhydroxylate analyte
and is generally "on" in the presence of bound polyhydroxylate analyte. Thus, the
reversible binding of a polyhydroxylate analyte essentially turns the fluorescent
switch "on" and "off". This property of the biosensor molecules, as well as their
ability to emit fluorescent light at greater than about 500 nm, renders these biosensor
molecules particularly well-suited for detecting and measuring in-vivo glucose concentrations.