Using
dissolved O2 as the cathodic co-reactant of three-dimensional
(3D) g-C3N4 is a convenient method to improve
the electrochemiluminescence (ECL) signal, but it still suffers the
disadvantages of limited luminous efficiency of 3D g-C3N4 and low content, low reactivity, and instability of
dissolved O2. Here, N vacancy with high density was first
introduced into the structure of 3D g-C3N4 (3D
g-C3N4-NV), which could conveniently realize
multipath ECL improvement by simultaneously solving the above shortcomings
effectively. Specifically, N vacancy could change the electronic structure
of 3D g-C3N4 to broaden its band gap, increase
fluorescence (FL) lifetime, and accelerate electron transfer rate,
obviously improving the luminous efficiency of 3D g-C3N4. Meanwhile, N vacancy made the excitation potential of 3D
g-C3N4-NV to shift from −1.3 to −0.6
V, effectively weakening the electrode passivation. Moreover, the
adsorption capacity of 3D g-C3N4-NV was obviously
enhanced, which could make the dissolved O2 enrich around
3D g-C3N4-NV. And massive active NV sites of
3D g-C3N4-NV could promote O2 to
more efficiently convert to reactive oxygen species (ROS) that were
key intermediates in ECL generation. Using the newly proposed 3D g-C3N4-NV-dissolved O2 system as an ECL
emitter, an ultrasensitive target conversion biosensor was constructed
for miRNA-222 detection. The fabricated ECL biosensor exhibited satisfactory
analytical performance for miRNA-222 with a detection limit of 16.6
aM. The strategy achieved multipath ECL improvement by introducing
high-density N vacancy simply in the 3D structure of g-C3N4 and could open a new horizon for developing a high-performance
ECL system.