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Intracellular viscosity plays an important role in regulating cellular morphology and physiology and is closely related to a host of diseases. Especially, the changes in mitochondrial viscosity will cause some common diseases such as hyperlipidemia, Alzheimer's disease and cancer. In this work, we report the design of a red-emissive molecular rotor for the detection of mitochondrial viscosity in live cells. The probe showed fascinating performance, such as specific targeting to mitochondria, high sensitivity to viscosity, and rapid fluorescence response, especially the dual response mode of fluorescence intensity and fluorescence lifetime. By using this probe, we realized monitoring of the mitochondrial viscosity variations in live cells under different physiological processes. Our study offers an opportunity to discover potential tools for mitochondria-related physiology and pathology investigation. [ABSTRACT FROM AUTHOR]

A ratiometric fluorescent probe with a large emission shift was developed for the accurate measurement of hydrogen peroxide (H2O2) in sophisticated pyroptosis signaling pathways. The results reported here demonstrate that H2O2, as a principal member of ROS, is a critical upstream signaling molecule in regulating pyroptosis. [ABSTRACT FROM AUTHOR]

Enzyme activity in live cells is dynamically regulated by small‐molecule transmitters for maintaining normal physiological functions. A few probes have been devised to measure intracellular enzyme activities by fluorescent imaging, but the study of the regulation of enzyme activity via gasotransmitters in situ remains a long‐standing challenge. Herein, we report a three‐channel imaging correlation by a single dual‐reactive fluorescent probe to measure the dependence of phosphatase activity on the H2S level in cells. The two sites of the probe reactive to H2S and phosphatase individually produce blue and green fluorescent responses, respectively, and resonance energy transfer can be triggered by their coexistence. Fluorescent analysis based on the three‐channel imaging correlation shows that cells have an ideal level of H2S to promote phosphatase activity up to its maximum. Significantly, a slight deviation from this H2S level leads to a sharp decrease of phosphatase activity. The discovery further strengthens our understanding of the importance of H2S in cellular signaling and in various human diseases. Two for one: A dual‐reactive molecular probe has been devised to simultaneously measure intracellular H2S levels and phosphatase activity. Fluorescent analysis by three‐channel imaging correlation reveals that a slight deviation from the normal H2S level leads to a sharp decrease of phosphatase activity. [ABSTRACT FROM AUTHOR]

Abstract: A novel dansyl-based fluorescent probe was synthesized and characterized. It exhibits high selectivity and sensitivity towards Fe3+ ion. This fluorescent probe is photostable, water soluble and pH insensitive. The limit of detection is found to be 0.62μM. These properties make it a good fluorescent probe for Fe3+ ion detection in both chemical and biological systems. Spike recovery test confirms its practical application in tap water samples. [Copyright &y& Elsevier]

Reactive oxygen species (ROS) regulating the release of free zinc ions (Zn 2+ ) in cellular lysosome is closely related to various pathways of cellular signal transduction, such as inflammation and oxidative stress. Directly visualizing Zn 2+ release in lysosome is essential for in-depth understanding these physiological processes, and is still an atelic challenge. In this work, we successfully fabricate a lysosome-specific Zn 2+ fluorescent probe and achieve the visualization of ROS-induced Zn 2+ release in lysosome of inflammatory cells. The as-prepared probe combines a green fluorophore, an ionophore with five-dentate sites, and a morpholine as the lysosome-specific localization moiety. The fluorescence of the fluorophore in the free probe is suppressed by a photoinduced electron transfer (PET) process from nitrogen atoms in the ionophore. Upon the addition of Zn 2+ , the fluorescence can be promoted immediately, achieving the real-time detection. Meanwhile, the probe is sensitive and selective to Zn 2+ , which provides the capability to detect low-concentration of free Zn 2+ in lysosomes. Accordingly, the Zn 2+ release was clearly observed in lysosome with the increase of ROS levels when the inflammation occurred in living cells. [ABSTRACT FROM AUTHOR]

The intracellular pH value controls many aspects of cell metabolism and is considered as an integral part of many metabolic reactions by maintaining the structure and function of nearly all proteins. Detection of pH fluctuation covering the whole intracellular pH window including both neutral and acidic is essential but challenging and still needs more sensitive pH probes. We have synthesized a new near-infrared fluorescent and colorimetric probe based on cyanine dye, which can be used for monitoring the pH fluctuation. The probe has a good water solubility, excellent photostability, reversibility, low cytotoxicity and exhibits a good performance to detect pH fluctuation in both solution and live cells. Upon incubation with live cells, it responds sensitively to low pH value in cell medium by exhibiting bright red fluorescence. On the contrary, the fluorescence intensity of probe seems to be very weak in neutral cellular environment. The satisfactory results show that the probe was a highly sensitive pH probe in living cells and further demonstrate its promising biological applications. [ABSTRACT FROM AUTHOR]

Investigating the interactions between lipid droplets (LDs) and other organelles are greatly significant to the in-depth understand the physiology and pathology of LDs. Despite of commercial LDs probes available, the ER and other membrane structures are also stained due to their amphiphilicity, which limits their further applications. To address this issue, we herein report a highly hydrophobic fluorescent probe to specifically label intracellular LDs with high signal/noise ratio, avoiding undesirable staining of ER and thus achieving to real-time monitor the interactions between LDs and other organelles. The probe showed strong emission in lipid environments but no fluorescence in water and liposomes, enabling the probe with high signal/noise ratio. Using this probe, it was found that the contacts of LDs with ER and mitochondria clearly increased under the condition of starvation. • A highly hydrophobic fluorescent molecule was successfully synthesized, which can specifically target lipid droplets. • The signal-to-noise ratio of the synthesized fluorescent probe is much higher than that of commercial dyes. • The interactions of lipid droplets with mitochondria and endoplasmic reticulum were studied. [ABSTRACT FROM AUTHOR]

• A single-molecular probe specific to mitochondria was constructed by the linkage of 7-hydroxylcoumarin and rhodamine B. • The as-synthesized fluorescent probe without FRET was sensitive to pH change in the range of 3–9. • The probe with almost no cytotoxicity was suitable to track mitochondrial fusion, fission and autophagy. Tracking of mitochondrial dynamic processes including their morphology, biogenesis and elimination is of significance to deeply understand mitochondrial physiology and related diseases, which requires simultaneous and discriminative labeling of both mitochondria themselves and correlated downstream organelles. However, the combination of multiple fluorescent probes will bring the large invasive effect that seriously impacts the normal physiological processes of mitochondria within live cells. To address this issue, we herein construct a single molecular probe through the combination of two pH-sensitive fluorogens, hydroxyl-coumarin and rhodamine B, enabling the detection of intracellular microenvironmental pH variations. Importantly, the as-synthesized probe not only accumulates into mitochondria, indicated by cyan emission from hydroxyl-coumarin but also can be delivered to the downstream acidic autolysosomes upon the occurrence of mitophagy, indicated by red emission from rhodamine B. Taking advantage of a two-channel confocal microscopy strategy, we have achieved to real-time track the processes of mitochondrial replication, merge and autophagy. [ABSTRACT FROM AUTHOR]

Understanding lysosome-related physiology needs specific lysosome probes to track the biological processes of lysosome in living cells. Here, we report an azacyclo-modified fluorescent probe that has a large Stokes shift, good photostability and negligible cytotoxicity for highly specific labeling of lysosome and autolysosome in living cells. The probes with different kinds of azacyclo groups on parent dye dansyl are screened to show that dansyl-cycleanine (DNS-C) with four nitrogen atoms possesses the best lysosome-localized ability. And DNS-C as a universal tracker exhibits excellent ability for lysosome labeling in different cell lines with high overlap coefficients (≥0.90). Different from a commercially available LysoTracker, the Stokes shift of DNS-C up to 240 nm (λ ex/em = 330/570 nm), is much larger than that of LysoTracker ~20 nm (λ ex/em = 573/595 nm). More importantly, the fluorescence of DNS-C keeps still high brightness after a time-lapsed imaging for 40 min in living cells, implying its remarkable photostability for long-term tracking. In addition, DNS-C can also clearly image the autolysosome, a critical subcellular compartment, forming by the fusion of lysosome with autophagosome in autophagy. These results suggest the promising utility of our probe as a powerful tool to real-time trace physiological processes of lysosomes. Image 1 • The azacyclo-localizing fluorescent probe DNS-C exhibits high specificity for lysosome and autolysosome labeling. • The large Stocks shift ~240 nm can efficiently avoid unwanted self-reabsorption and inner-filter effects. • The DNS-C can serve as a universal tracker for lysosome labeling in different kinds of cells. • The DNS-C exhibits excellent photostablility for long-term tracking the lysosome-related behaviors. [ABSTRACT FROM AUTHOR]