Rational design of specific ratiometric viscosity probes with small molecular weight is a challenge in practical biotechnology applications. Herein two novel water-soluble, small-molecular ratiometric probes, bearing N-methyl benzothiazolium moiety (DSF and DBF), are designed for two-photon fluorescent imaging as a functional of local viscosity. The dye DSF, a light-up fluorescent probe, is sensitive to local viscosity and selectively stains nuclear DNA, which can be used to inspect asynchronous cells under confocal microscopy. While the dye DBF as a molecular rotor displays strong fluorescence enhancement in viscous media or binding to RNA. It exhibits dual absorption and emission as well, and only the red emission is markedly sensitive to viscosity changes, providing a ratiometric response and selectively imaging nucleolic and cytosolic RNA. Interestingly it is shown, for the first time, that the intracellular targeting and localization (DNA and RNA) of the two dyes are entirely realized simply by modifying the substituent attached to the benzothiazolium.

Photodynamic therapy (PDT), as an emerging treatment modality, which takes advantage of reactive oxygen species (ROS) generated upon light illumination to ablate tumours, has suffered from a limited treatment depth, strong oxygen dependence and short ROS lifespan. Herein, we developed a highly efficient NIR-I light (808 nm laser) initiated theranostic system based on a fluorescent photosensitizer (EBD-1) with cancer cell membrane targeting ability, which can realize large penetration depth in tissue, generate superoxide radicals (O₂ ⁻˙) to relieve the oxygen-dependence, confine the ROS oxidation at the cell membrane, and self-report the cell viability during the PDT process. In vivo experiments demonstrated that EBD-1 under 808 nm light successfully accomplished remarkable cancer ablation. This work will be beneficial for the design of novel photosensitizers for PDT-based theranostic systems.

We present a two-photon (2P, 800 nm) PDT cyclometalated Iridium(III) complex (Ir-Es) that targets the intracellular nucleus. The complex is capable of migrating sequentially from the nucleus to mitochondria and inducing dual-damage under light exposure. This study suggests that with minor modification of the terminal moieties of complexes, their final intracellular destinations and PDT efficiency can be significantly impacted.

Multiphoton bioimaging benefits from good penetration of tissue, low phototoxicity and high resolution. Hence, development of efficient multiphoton imaging agents is highly desirable but remains challenging. Herein, a novel terpyridine-based Zn(II) complex bearing a thiophene bridge was designed rationally and fabricated. Thanks to its aggregation-induced emission (AIE), DZ1 emitted bright yellow-green fluorescence (λ_em = 575 nm) under physiological conditions. The three-photon spectral changes of DZ1 when binding with RNA unambiguously reflected its RNA-specific targeting behaviour, resulting in twofold enhancement in three-photon action cross-sections located at the second near-infrared window (1700 nm).