Fluorescence tunable carbon dots for in vitro nuclear dynamics and gastrointestinal imaging in live zebrafish and their in vivo toxicity evaluation by cardio-craniofacial disfunction assessment

Abstract

Sub-cellular organelle anomalies are frequently observed in diseases such as cancer. Early and precise diagnosis of these alterations can be crucial for patient outcomes. However, current diagnostic tools using conventional organic dyes or metal quantum dots face limitations, including poor biocompatibility, stringent storage conditions, limited solubility in aqueous media, and slow staining speeds. These challenges underscore the need for safer, more effective diagnostic and therapeutic solutions. In these aspects, we have developed highly photostable, biocompatible, water-dispersible carbon dots (TNCDs) with an average size of 5.5 nm using tartaric acid and ethylenediamine via a hydrothermal route. The synthesized TNCDs have shown bright blue fluorescence under the irradiation of UV-light at an excitation wavelength of 365 nm. They exhibit a quantum yield (QY) of 25.1% with maximum emission at 390 nm. A nice tri-exponential fitting of the decay curve with characteristic lifetimes of 1.52 ns, 3.05 ns and 6.11 ns for TNCDs was obtained. In vitro studies demonstrated that TNCDs have high biocompatibility (20 μg ml⁻¹) with almost 100% cell viability and excellent nucleus targeting and staining capabilities with low background interference (with 10–12 times enhancement in fluorescence intensity). Additionally, if tagged with photosensitizers or radionuclides, TNCDs can serve as therapeutic agents in photodynamic therapy against cancer cells. Importantly, TNCDs exhibited negligible toxicity in developing zebrafish even at high concentrations (up to 400 mg L⁻¹) as investigated by cardio and craniofacial disfunction assessment. Live organism imaging revealed that TNCDs produced aggregation-induced strong and specific green fluorescence in the gut of zebrafish larvae even at low concentrations, indicating their potential for nucleus staining and gut-specific optical imaging (at 50 mg L⁻¹). Thus, our TNCDs represent a robust nanoplatform for cellular and whole-organism fluorescence imaging, offering both diagnostic and therapeutic potential.