RESUMO
In situ visualization of lipid composition diversity in lipid droplets (LDs) is essential for decoding lipid metabolism and function. However, effective probes for simultaneously localizing and reflecting the lipid composition of LDs are currently lacking. Here, we synthesized full-color bifunctional carbon dots (CDs) that can target LDs as well as respond to the nuance in internal lipid compositions with highly sensitive fluorescence signals, due to lipophilicity and surface state luminescence. Combined with microscopic imaging, uniform manifold approximation and projection, and sensor array concept, the capacity of cells to produce and maintain LD subgroups with varying lipid composition was clarified. Moreover, in oxidative stress cells, LDs with characteristic lipid compositions were deployed around mitochondria, and the proportion of LD subgroups changed, which gradually disappeared when treated with oxidative stress therapeutics. The CDs demonstrate great potential for in situ investigation of the LD subgroups and metabolic regulations.
Assuntos
Gotículas Lipídicas , Mitocôndrias , Gotículas Lipídicas/metabolismo , Mitocôndrias/metabolismo , Metabolismo dos Lipídeos , LipídeosRESUMO
Lysosomal acidity relies on H+ inflow, which requires counter-ion flows (Cl- and K+) to balance charge. A lysosome targeting ternary recognition fluorescent probe for Cl-, K+, and pH was developed for lysosome acidification counter-ion research. The probe was used to study counter-ion changes when the Cl- channel was blocked and under oxidative pressure.
RESUMO
TMR-TPE, a ratiometric fluorescent probe, was reported for the imaging of vicinal dithiol proteins (VDPs) in living cells. Profiting from the "dual-key-and-lock" design, TMR-TPE solves the toxicity problem of VDP probes (98% cell viability at 50 µM) and avoids the interference of small thiols (up to 10 mM GSH). The change of VDPs during drug-induced liver injury was monitored for the first time using TMR-TPE.
Assuntos
Corantes Fluorescentes , Proteínas , Corantes Fluorescentes/toxicidade , Corantes Fluorescentes/metabolismo , Proteínas/metabolismo , Microscopia de Fluorescência/métodos , Compostos de SulfidrilaRESUMO
Endogenous formaldehyde (FA) exists in many living cells and in inhomogeneous distribution in organelles. In particular, lysosomes play significant roles in FA generation and the biofunction of living cells. Herein, we developed a new ratiometric fluorescent nanoprobe, based on naphthalimide derivative (ND)-functionalized carbon dots (CDs), for monitoring endogenous FA in lysosomes. The fluorescence intensity (F535) of green-emitting ND at 535 nm serves as the response signal and the fluorescence intensity (F414) of blue-emitting CDs at 414 nm acts as the reference signal. The fluorescence intensity ratio (F535/F414) of the CD-ND probe is linearly correlated with FA concentration within the range of 1-40 µM in aqueous solution, and the detection limit (3σ/slope) is estimated to be 0.34 µM. As for practical application, this nanoprobe is utilized for the ratiometric fluorescence imaging of FA in live cells. Remarkably, this nanoprobe can specifically target and stain the lysosomes and detect exogenous and endogenous FA in HeLa cells. The new FA probe shows a superior lysosomal targeting ability with a Pearson's coefficient of 0.93, which is attributed to the macromolecular size and basic amine group functionalized surface of CD-ND.
Assuntos
Carbono/química , Corantes Fluorescentes/química , Formaldeído/análise , Lisossomos/metabolismo , Microscopia Confocal , Naftalimidas/química , Pontos Quânticos/química , Clorpromazina/farmacologia , Endocitose/efeitos dos fármacos , Células HeLa , Humanos , Limite de Detecção , Pontos Quânticos/metabolismo , Espectrometria de FluorescênciaRESUMO
Great efforts have been made to develop facile and efficient methods to prepare carbonaceous nanostructures with long-wavelength emission. Herein, we report a low-temperature aqueous strategy to synthesize red-emissive carbon polymer dots (R-CPDs) through the regulation of oxidative polymerization of p-phenylenediamine at 80 °C. The morphology, chemical composition and photophysical properties of the R-CPDs are characterized and analyzed in detail, thereby elucidating their photoluminescence origins from the surface state and crosslink enhanced emission effect. The resulting R-CPDs possess unique features including high pH-sensitivity within pH 4-6 and a wide-range tunable solvent-color effect (λem 528-600 nm). Moreover, the R-CPDs show high stability in physiological media with high salinity, and good resistance to photobleaching. In addition to their favorable biocompatibility, the R-CPDs are applied for monitoring the pH fluctuation in HeLa cells. This study not only provides a unique red emissive carbonaceous nanomaterial for cellular imaging and multicolor applications, but also presents a novel perspective for the construction of long-wavelength emission carbon-based nanomaterials by simple and controllable strategies.