Dual-Mode Gold Nanocluster-Based Nanoprobe Platform for Two-Photon Fluorescence Imaging and Fluorescence Lifetime Imaging of Intracellular Endogenous miRNA

Bioimaging is widely used in various fields of modern medicine. Fluorescence imaging has the advantages of high sensitivity, high selectivity, noninvasiveness, in situ imaging, and so on. However, one-photon (OP) fluorescence imaging has problems, such as low tissue penetration depth and low spatiotemporal resolution. These disadvantages can be solved by two-photon (TP) fluorescence imaging. However, TP imaging still uses fluorescence intensity as a signal. The complexity of organisms will inevitably affect the change of fluorescence intensity, cause false-positive signals, and affect the accuracy of the results obtained. Fluorescence lifetime imaging (FLIM) is different from other kinds of fluorescence imaging, which is an intrinsic property of the material and independent of the material concentration and fluorescence intensity. FLIM can effectively avoid the fluctuation of TP imaging based on fluorescence intensity and the interference of autofluorescence. Therefore, based on silica-coated gold nanoclusters (AuNCs@SiO2) combined with nucleic acid probes, the dual-mode nanoprobe platform was constructed for TP and FLIM imaging of intracellular endogenous miRNA-21 for the first time. First, the dual-mode nanoprobe used a dual fluorescence quencher of BHQ2 and graphene oxide (GO), which has a high signal-to-noise ratio and anti-interference. Second, the dual-mode nanoprobe can detect miR-21 with high sensitivity and selectivity in vitro, with a detection limit of 0.91 nM. Finally, the dual-mode nanoprobes performed satisfactory TP fluorescence imaging (330.0 mu m penetration depth) and FLIM (tau(ave) = 50.0 ns) of endogenous miR-21 in living cells and tissues. The dual-mode platforms have promising applications in miRNA-based early detection and therapy and hold much promise for improving clinical efficacy.

Automated summary

Bioimaging, including fluorescence imaging, is widely used in modern medicine. However, one-photon fluorescence imaging has limitations such as low tissue penetration depth and low spatiotemporal resolution. Two-photon fluorescence imaging can overcome these limitations, but the complexity of organisms can affect the accuracy of results. Fluorescence lifetime imaging (FLIM) is an intrinsic property of materials and independent of material concentration and fluorescence intensity, making it a promising alternative. By combining silica-coated gold nanoclusters (AuNCs@SiO2) with nucleic acid probes, a dual-mode nanoprobe platform for two-photon and FLIM imaging of intracellular miRNA-21 was constructed. The dual-mode nanoprobe exhibited high sensitivity and selectivity, with potential applications in miRNA-based early detection and therapy.