A mitochondria-targeted single fluorescence probe for separately and continuously visualizing H2S and Cys with multi-response signals.

Hydrogen sulfide and cysteine are momentous endogenous regulators of many physiological processes and maintain a dynamic balance of redox in living organisms. To investigate the inter-relationship of them in vivo, there is a pressing need to develop analytical molecular tools to identify related biomolecules. We construct a mitochondria-targeted single fluorescence probe (Mit-CM) for separately and continuously visualizing H2S, Cys and H2S/Cys with multi-response fluorescence signals. Mit-CM has the following advantages: (Ⅰ) colorimetric and ratiometric: two well-separated emission bands can ensure accurate detection of the analyte and significant color changes contribute to rapid identification of the analyte by the naked eye; (Ⅱ) mitochondrial localization: study the physiological functions of H2S and Cys in mitochondria; (Ⅲ) separate and continuous detection of H2S and Cys: reveal the inter-relationship and interconversion of them in biological system. Moreover, the desirable attributes of low cytotoxicity, better biocompatibility and excellent mitochondria enrichment ability indicate that Mit-CM can be employed to achieve detection and observe distribution of H2S, Cys and H2S/Cys in living organism.

A novel fluorescent probe for rapidly detection cysteine in cystinuria urine, living cancer/normal cells and BALB/c nude mice.

Cysteine (Cys), an important organic small molecule containing sulfhydryl groups, plays paramount functions in human pathologies and physiologies. The detection of Cys in living vivo is essential for studying its roles. Here, we designed and synthesized a novel red-emission fluorescent probe AXPI-Cys with highly sensitivity (LOD = 48.9 ±â€¯0.23 nM), rapidly response (<7 min) and colorimetric for detection cysteine. More importantly, the AXPI-Cys was determined Cys in real cystinuria urine samples for the first time with the satisfactory results (92%-99.96%) and employed for specifically location of endogenous/exogenous Cys in living cancer/normal cells and almost non-toxic, that is very valuable for diagnosis of cystinuria and observation of the distribution of Cys in normal cells. Notably, the AXPI-Cys was applied to imaging Cys in BALB/c nude mice with good biocompatibility and desirable tissue-penetration depth. Owing to the superior capability of AXPI-Cys, it provided a desired method to detect Cys in urine samples and cells, and exhibited munificent potential usage in biosystems and imaging studies in vivo.

Dual-Functional NIR AIEgens for High-Fidelity Imaging of Lysosomes in Cells and Photodynamic Therapy.

Design and synthesis of water-soluble near-infrared (NIR) emissive fluorescent molecules with aggregation-induced emission (AIE) characteristics, perfect signal-to-noise ratio for imaging of organelle, and photodynamic therapy (PDT) functions has received enormous attention. However, the dual-functional NIR AIEgens of high-fidelity tracking lysosome and ablation cancer cells was rarely reported. Herein, a series of AIE luminogens (AIEgens) with a typical AIE effect, good biocompatibility, superior photostability, high brightness, and excellent reactive oxygen species (ROS) generation ability were developed, which had different electronic push-pull strength and conjugate system size in the molecular structure. These AIEgens could specifically "light up" and dynamically long-term track the lysosomes in living cells and zebrafish with ultrahigh colocalization imaging Pearson's correlation coefficients (Rr: 0.9687) and overlap coefficient (R: 0.9967). Additionally, the MPAT of NIR luminescence as a photosensitizer was used for photodynamic ablation of cancer cells, owing to prompt generation of the ROS under green light irradiation (495-530 nm, 10 mW cm-2). Hence, this research not only expands the application range of NIR AIEgens but also provides useful insights into design of split-new method for the treatment of cancer.

A highly selective ratiometric fluorescent probe based on naphthalimide for detection and imaging of CYP1A1 in living cells and zebrafish.

Real-time monitoring of the cytochrome P450 1A1 (CYP1A1) activity in complex biological systems via a practical tool is highly sought after because of its significant role in the metabolism and bioactivation of various xenobiotics. Herein, according to slight differences in the 3D structure and substrate preference between CYP1A1 and its homologous CYP1A2, a series of novel ratiometric fluorescent probes were designed and synthesized using 1,8-naphthalimide because of its trait of naked-eye visualization and ratiometric fluorescence to achieve the detection of CYP1A1 in biological samples. Among these probes, NEiPN showed good water solubility, highly isoform selectivity and great sensitivity (LOD = 0.04874 nM) for CYP1A1 under simulated physiological conditions, which makes it favorable for monitoring CYP1A1 in vivo. Remarkably, NEiPN exhibited excellent reproducibility when it was used to detect the CYP1A1 content in human liver microsomes, which indicated that it has a great potential for quantifying the CYP1A1 content in real biological samples. Furthermore, NEiPN showed relatively low cytotoxicity and has been successfully applied in biological imaging in living cells and zebrafish. These findings indicate that NEiPN is capable of real-time monitoring of the activity of endogenous CYP1A1, which could provide support for CYP1A1-associated pathological processes.

A novel probe for colorimetric and near-infrared fluorescence detection of cysteine in aqueous solution, cells and zebrafish.

Cysteine(Cys) is tightly related to physiological and pathological of human, and the imbalance of concentration of cysteine in the intracellular are associated with many diseases. Here, a novel NIR fluorescent probe TCF-Cys was designed and synthesized, and both the optimal excitation and emission wavelength of them were between 650 and 900 nm, that within the "optical window" of biological tissues. In aqueous solution, TCF-Cys, which with an acrylate extremity as a recognizing unit, exhibited excellent "turn-on" fluorescence response for Cys superior to other amino acids and thiols with a limit of detection of 0.1323 µM. Moreover, as an excellent naked-eye colorimetric indicator, TCF-Cys could effectively distinguishing the Cys, Hcy and GSH in aqueous solution through color change. Then, the response mechanism of TCF-Cys for Cys was revealed by TLC, 1H NMR, HPLC, HRMS and DFT calculation. Finally, TCF-Cys was successfully employed to fluorescence specifically map of exogenous and endogenous Cys in living cells and zebrafish with low toxicity.