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Lysosome-Targeted Single Fluorescence Probe for Two-Channel Imaging Intracellular SO2 and Biothiols
摘要： As the members of reactive sulfur species, SO2 and biothiols play a signi?cant role in physiological and pathological processes and directly in?uence numerous diseases. Furthermore, SO2 and biothiols can provide a reductive environment for lysosomes to carry out their optimal functionality. To this end, the development of single ?uorescent probes for imaging SO2 and biothiols from different emission channels is highly desirable for understanding their physiological nature. Here, a lysosome-targeted ?uorescent probe (BPO-DNSP) with a dual reaction site for SO2 and biothiols was presented. BPO-DNSP can sensitively and selectively respond to SO2 in the green channel with a large Stokes shift over 105 nm, and to biothiols in the near-infrared emission channel with a large Stokes shift over 109 nm. The emission shift for the two channels was as high as 170 nm. Colocalization experiments veri?ed that BPO-DNSP can selectively enrich lysosomes. Notably, BPO-DNSP can not only be used to image intracellular SO2 and biothiols from two different channels, but also to monitor the conversion of biothiols to SO2 without adding exogenous enzymes in living HeLa cells.
关键词： ?uorescence imaging,lysosome-targeted,single ?uorescent probe,biothiols,SO2
更新于2025-11-21 11:08:12
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Intriguing “chameleon” fluorescent bioprobes for the visualization of lipid droplet-lysosome interplay
摘要： The interplay of lipid droplets (LDs) and lysosome plays an important role in cell metabolism, and the visualization of this process can provide useful information of organelle communication and function. However, fluorescent bioprobes based on organic fluorophores that can respond to LD-lysosome interplay are much rare. Herein, fluorescent bioprobes with high photostability, excellent biocompatibility and intracellular polarity sensitivity are achieved by encapsulating a new red fluorogenic molecule TPA-BTTDO within polymeric matrix (DSPE-PEG2000). They can sequentially localize in lysosome and LDs with red and cyan emissions, respectively. By monitoring the emission color change, the interesting dynamic processes of the probes escaping from lysosome and then enriching in LDs, and finally returning to lysosome after LDs consumption are visualized. In addition, the tracing of dynamic movement and consumption of LDs is realized by the probes with a high signal-to-noise ratio. The unique labeling behaviors and distinguished dual emissions of the probes in LDs and lysosome make them promising agents for fluorescence visualization studies of LD-lysosome related bioprocess and metabolism diseases.
关键词： lipid droplet,aggregation-induced emission,nanomaterials,lysosome,fluorescent bioprobe
更新于2025-11-21 11:08:12
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Ratiometric fluorescent probe based on pyrrole-modified rhodamine 6G hydrazone for the imaging of Cu2+ in lysosomes
摘要： A novel rhodamine-based Schiff base derivative was obtained via the simple condensation of substituted formyl-1H-pyrrole and rhodamine 6G hydrazone. Fluorescence resonance energy transfer enabled the subsequent use of the derivative as a naked-eye colorimetric and ratiometric fluorescent sensor for Cu2+ in semi-aqueous solution, and the existence of the morpholine group enabled the further application of the sensor in imaging Cu2+ in the lysosomes of HeLa cells.
关键词： ratiometric fluorescent probe,lysosome-targeting,rhodamine hydrazone,Cu2+,FRET,pyrrole
更新于2025-09-23 15:23:52
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A new lysosome-targetable fluorescent probe with a large Stokes shift for detection of endogenous hydrogen polysulfides in living cells
摘要： Hydrogen polysulfides (H2Sn, n>1) influence a variety of important biological functions and activities associated with hydrogen sulfide (H2S). The development of probes for rapid, selective, and sensitive detection of H2Sn still remains a great challenge. Lysosome plays crucial roles in various physiological processes among living cells, which arouse high interest in detecting endogenous lysosome-targetable H2Sn. To the best of our knowledge, there is no lysosome-targetable probe for H2Sn has been reported. In this work, a new lysosome-targetable probe NIPY-NF, based on the scaffold of imidazo[1,5-α]pyridine, with a large Stokes shift (215 nm), low detection limit (84 nM), fast response time (6 min) and low cytotoxicity was designed and synthesized. Furthermore, NIPY-NF was successfully applied into the cell imaging for detection of endogenous lysosome-targetable H2Sn.
关键词： Hydrogen polysulfides,Fluorescent probe,Lysosome,Cell imaging,Imidazo[1,5-α]pyridine
更新于2025-09-23 15:23:52
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A pH-insensitive near-infrared fluorescent probe for wash-free lysosome-specific tracking with long time during physiological and pathological processes
摘要： A novel dicyanomethylene-4H-pyran based lysosome-specific near-infrared (NIR) probe, DCM-ML, is reported. It exhibits very weak fluorescence (Ф=0.007) in aqueous solution due to the energy relaxation from intramolecular rotation; but a large "off-on" NIR emission (20-folds) after specifically labeling lysosomes due to the local high viscosity instead of the local pH by both one- and two-photon excitation microscopy. Therefore, the imaging ability of DCM-ML will not confront the problems suffered by the currently used and reported pH-dependent lysosome probes (e.g., once lysosomal pH increases, their fluorescence gets quenched); The pH-independent feature of DCM-ML is further demonstrated to be a great advantage for tracing lysosomes’ movements in a relatively long time, even during cellular processes when lysosomes are suffered from external stimulations that induce pH increase and apoptosis. In addition to the characteristics of pH-insensitivity and "off-on" NIR emission, DCM-ML also possesses characteristics of excellent membrane permeability, high selectivity, low cytotoxicity, good photostability and large Stokes shift (> 200 nm), which qualify it as a superior wash-free NIR probe for tracking dynamic changes of lysosomes under both physiological and toxicological conditions. Hence, we believe DCM-ML may find a great potential of application in the future.
关键词： pH-insensitivity,Long-term tracking,Wash-free,"Off-on",Lysosome-specific imaging,Near-infrared
更新于2025-09-23 15:22:29
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Fluorescent half-sandwich phosphine-sulfonate iridium(III) and ruthenium(II) complexes as potential lysosome-targeted anticancer agents
摘要： The synthesis, characterization and biological activity of neutral fluorescent Ir(III) and Ru(II) half-sandwich organometallic complexes containing phosphine-sulfonate ligands are reported. X-ray crystal structure of complexes 1-3, 10 and 11 exhibits the expected half-sandwich “three-legged piano-stool” pseudo-octahedral geometry. Spectroscopic properties study displays that these complexes show rich fluorescence properties. With the exception of 9, 10 and 11 toward A549 human lung cancer cells and 10 towards HeLa human cervical cancer cells, each complex shows promising cytotoxicity toward HeLa and A549 cells line with IC50 values in the range of 3.6-53.1 μM, and 6.5-34.5 μM, respectively. Hydrolysis, DNA cleavage and depolarization of the mitochondrial membrane potential (MMP) appear not to be the main mechanism of action. However, these complexes are able to covert NADH to NAD+ via the transfer hydrogenation. Mechanism studies by flow cytometry display that the complexes exert their anticancer efficacy by inducing apoptosis, perturbing the cell cycle and increasing the intracellular ROS level. Furthermore, fluorescence property of these complexes provides a tool to investigate the microscopic mechanism by confocal microscopy. Notably, the typical Ir(III) complex 3 can specially localize to lysosome and damage it. In addition, complex 3 enters into HeLa cells mainly through energy-dependent pathway.
关键词： phosphine-sulfonate,half-sandwich,fluorescent,anticancer complexes,lysosome-targeted
更新于2025-09-23 15:21:21
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Blue light-emitting diode irradiation promotes transcription factor EB-mediated lysosome biogenesis and lysosomal cell death in murine photoreceptor-derived cells
摘要： Exposure to blue light from light-emitting diodes (LEDs) is a source of damage for human eyes in today’s modern life. Although it is well known that blue light can cause cellular damage and death, the molecular mechanism underlying this is still not fully understood. Here, we demonstrated that exposure to blue LED light increased lysosome levels and perinuclear cluster formation in 661W murine photoreceptor-derived cells. Irradiation with blue LED light promoted the nuclear transport of transcription factor EB (TFEB) and a subsequent increase in lysosomal-related gene expression. Moreover, blue LED light induced morphological changes in lysosomal structure and lysosomal membrane permeabilization (LMP). These effects were suppressed by an antioxidant, N-acetylcysteine (NAC). Finally, a calcium ion chelator, BAPTA-AM, attenuated blue LED light-induced lysosomal biogenesis and cell death. Taken together, these findings suggest that oxidative stress under blue LED light increases lysosome levels via the TFEB pathway in a calcium-dependent manner, resulting in the accumulation of damaged lysosomes and subsequently lysosomal cell death. Our results imply that lysosomal homeostasis plays a key role in the maintenance of eye function and the progression of retinal diseases.
关键词： TFEB,Blue LED light,Calcium,Lysosome biogenesis,Oxidative stress,Lysosomal membrane permeabilization
更新于2025-09-23 15:21:01
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<p>A Lysosome-Targetable Fluorescence Probe Based on L-Cysteine-Polyamine-Morpholine-Modified Quantum Dots for Imaging in Living Cells</p>
摘要： Quantum dots (QDs) are used as fluorescent probes due to their high fluorescence intensity, longevity of fluorescence, strong light-resistant bleaching ability and high light stability. Therefore, we explore a more precise probe that can target an organelle. Methods: In the current study, a new class of fluorescence probes were developed using QDs capped with 4 different L-cysteine-polyamine-morpholine linked by mercapto groups. Ligands were characterised by Electrospray ionization mass spectrometry (ESI-MS), H-Nuclear Magnetic Resonance (1H NMR) spectroscopy, and 13C NMR spectroscopy. Modified QDs were characterized by Transmission Electron Microscope (TEM), Ultraviolet and visible spectrophotometry (UV–Vis), and fluorescence microscopy. And the biological activity of modified QDs was explored by using MTT assay with HeLa, SMMC-7721 and HepG2 cells. The fluorescence imaging of modified QDs was obtained by confocal laser scanning fluorescence microscopy (CLSM). Results: Synthesized QDs ranged between 4 to 5 nm and had strong optical emission properties. UV–Vis and fluorescence spectra demonstrated that the cysteine-polyamine-morpholine were successfully incorporated into QD nanoparticles. The MTT results demonstrated that modified QDs had lesser cytotoxicity when compared to unmodified QDs. In addition, modified QDs had strong fluorescence intensity in HeLa cells and targeted lysosomes of HeLa cells. Conclusion: This study demonstrates the modified QDs efficiently entered cells and could be used as a potential lysosome-targeting fluorescent probe.
关键词： lysosome-targetable,cells imaging,quantum dots,fluorescence probe
更新于2025-09-23 15:19:57
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Multicolor Flow Cytometry-based Quantification of Mitochondria and Lysosomes in T Cells
摘要： T cells utilize different metabolic programs to match their functional needs during differentiation and proliferation. Mitochondria are crucial cellular components responsible for supplying cell energy; however, excess mitochondria also produce reactive oxygen species (ROS) that could cause cell death. Therefore, the number of mitochondria must constantly be adjusted to fit the needs of the cells. This dynamic regulation is achieved in part through the function of lysosomes that remove surplus/damaged organelles and macromolecules. Hence, cellular mitochondrial and lysosomal contents are key indicators to evaluate the metabolic adjustment of cells. With the development of probes for organelles, well-characterized lysosome or mitochondria-specific dyes have become available in various formats to label cellular lysosomes and mitochondria. Multicolor flow cytometry is a common tool to profile cell phenotypes, and has the capability to be integrated with other assays. Here, we present a detailed protocol of how to combine organelle-specific dyes with surface markers staining to measure the amount of lysosomes and mitochondria in different T cell populations on a flow cytometer.
关键词： lysosome,Flow cytometry,organelle-specific dyes,mitochondria,multicolor,T cell
更新于2025-09-19 17:15:36
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Instantaneous fluorescent probe for the specific detection of H2S
摘要： Novel cyanine-based fluorescent probes for the detection of H2S were developed. The probes developed are stable under physiological conditions. The water soluble fluorescent probe 2 displayed ultrafast and specific response to H2S displaying NIR fluorescence of 115-fold turn-on with the detection limit of 11 nM without assistance of organic solvent or surfactant. Cell imaging experiments indicated that probe 2 was cell-permeable and was able to detect H2S sensitively in lysosomes. Moreover, our probe was able to detect intrinsically produced H2S through enzymatic/non-enzymatic biosynthetic pathway from Cys/GSH. Moreover, we applied probe 2 to detect H2S in living mice and demonstrated the fast metabolism of H2S. Thus, probe 2 shows great promise as a reporter for H2S.
关键词： lysosome-targeted,H2S,fluorescent probe
更新于2025-09-19 17:15:36