Lanthanide-doped
upconversion nanoparticles (UCNPs) as energy donors
for Förster resonance energy transfer (FRET) are promising
in biosensing, bioimaging, and therapeutic applications. However,
traditional FRET-based UC nanoprobes show low efficiency and poor
sensitivity because only partial activators in UCNPs possessing suitable
distance with energy acceptors (<10 nm) can activate the FRET process.
Herein, a novel excited-state energy distribution-modulated upconversion
nanostructure is explored for highly efficient FRET. Integration of
the optimal 4% Er3+ doped shell and 100% Yb3+ core achieves ∼4.5-fold UC enhancement compared with commonly
used NaYF4:20%Yb3+,2%Er3+ nanoparticles,
enabling maximum donation of excitation energy to an acceptor. The
spatial confinement strategy shortens significantly the energy-transfer
distance (∼4.5 nm) and thus demonstrates experimentally a 91.9%
FRET efficiency inside the neutral red (NR)-conjugated NaYbF4@NaYF4:20%Yb3+,4%Er3+ nanoprobe,
which greatly outperforms the NaYbF4@NaYF4:20%Yb3+,4%Er3+@SiO2@NR nanoprobe (27.7% efficiency).
Theoretical FRET efficiency calculation and in situ single-nanoparticle
FRET measurement further confirm the excellent energy-transfer behavior.
The well-designed nanoprobe shows a much lower detection limit of
0.6 ng/mL and higher sensitivity and is superior to the reported NO2
– probes. Our work provides a feasible strategy
to exploit highly efficient FRET-based luminescence nanoprobes for
ultrasensitive detection of analytes.