Dopamine
and polyethyleneimine (PEI) copolymerized nanodots (PDA–PEI
nanodots) with both fluorescence emission and quenching features were
synthesized by a simple one-step reaction at room temperature. By
adjusting the dopamine and PEI ratio as well as the chain length of
PEI, the fluorescence emission and quenching properties of PDA–PEI
nanodots can be controlled well. Under optimal conditions, the nanodots
showed strong green fluorescence emission with an absolute quantum
yield of 1–2% and a quenching efficiency of more than 99% to
several fluorophores with emission wavelengths ranging from blue to
red light regions. The nanodots with a large number of functional
groups also showed strong affinity to nucleic acid strands, excellent
solubility in aqueous solution, long-term stability, and uniform size
distribution. Integrating these attractive features with the specific
enzymatic digestion reaction of the DSN enzyme, a highly sensitive
ratiometric fluorescence nanoprobe for miRNA analysis was developed.
Aminomethylcoumarin acetate (AMCA), which possesses the same excitation
wavelength but a well-resolved blue fluorescence emission with PDA–PEI
nanodots, was selected as the signal-reporting unit for capture probe
labeling, while the inherent green fluorescence of PDA–PEI
nanodots served as the reference. According to the ratiometric fluorescence
signal, the ratiometric fluorescence nanoprobes showed high sensitivity
and good accuracy for the miRNA assay. Because of the high and universal
quenching efficiency, stable fluorescence emission, easily assembled
interface, and uniform morphology, the nanodots may have great application
prospects to serve as a universal nanoplatform for the fabrication
of ratiometric fluorescence nanoprobes.