RESUMEN
Lipid droplets (LDs) and lysosomes play key roles in autophagy and cell apoptosis, and the discriminative visualization of the two organelles and simultaneously of autophagy and apoptosis is very helpful to understand their internal relationships. However, fluorescent probes that can concurrently achieve these tasks are not available currently. Herein, we delicately fabricate a robust probe CAQ2 for multiple tasks: illumination of LDs and lysosomes in dual emission colors as well as discriminative visualization of cell apoptosis and autophagy. The probe exhibited both lipophilic and basic properties and displayed different emission colors in neutral and protonated forms; thus, LDs and lysosomes emitted blue and red fluorescence colors, respectively. Because of the lysosomal acidification during autophagy, CAQ2 detected autophagy with evidently enhanced red emission. Because of the lysosomal alkalization during apoptosis, CAQ2 imaged apoptosis with a drastically decreased red fluorescence intensity. With the robust probe, the autophagy under starvation and lipidless conditions was visualized, and the apoptosis induced by H2O2, ultraviolet (UV) irradiation, and rotenone treatment was successfully observed. The efficient detoxification of Na2S against rotenone treatment was successfully revealed.
Asunto(s)
Colorantes Fluorescentes , Gotas Lipídicas , Peróxido de Hidrógeno , Rotenona , Lisosomas , Apoptosis , AutofagiaRESUMEN
Biomembranes in the endoplasmic reticulum (ER) play indispensable roles in various bioactivities, and therefore, visualizing the phase separation in ER membranes is crucial for the studies on the fundamental biology of the ER. However, near-infrared (NIR) ratiometric imaging of the phase behaviors of the ER in living cells with different statuses and in diverse tissues has not been investigated. Herein, we developed a polarity-responsive NIR fluorescent probe (DCA) for the visualization of the phase behavior in ER membranes. The probe displayed a large Stokes shift and was highly sensitive to polarity. By direct and native fluorescence imaging at room temperature, the ERo and ERd biomembranes in the ER could be clearly distinguished by dual NIR emission colors. Oxidative damage by H2O2 and homocystein (Hcy)-induced ER stress can efficiently induce the formation of large-scale ERo domains in ER membranes. Moreover, we have also revealed that different tissues exhibited diverse phase behaviors in the ER membranes. The ER membranes in cardiac and skeletal muscle tissues showed no evident phase separation, while large-scale ERo domains existed in the ER of liver tissues and formed at the ER membranes adjacent to lipid droplets (LDs) in white adipose tissues. We expect that the probe could serve as a powerful molecular tool to promote fundamental research studies on ER membranes and relative biomedical areas.
Asunto(s)
Peróxido de Hidrógeno , Imagen Óptica , Retículo Endoplásmico , Colorantes Fluorescentes , Gotas Lipídicas , Imagen Óptica/métodosRESUMEN
The in-depth study of the interplay and cooperation between multiple organelles is an important biological task. Single fluorescent probes for separate visualization of multiple organelles is a desirable molecular tool, but the construction of such a probe is extremely difficult owing to the lack of valid strategies. In this work, utilizing the reversible cyclization reaction and intermolecular π stacking mechanism, a robust fluorescent probe is constructed to discriminatively illuminate lipid droplets (LDs), mitochondria, and lysosomes with blue, green, and red emission colors, respectively. Using the probe, the interplays and cooperation between LDs, mitochondria, and lysosomes are successfully studied, and the critical roles of lysosomes and LDs during mitochondrial fission are successfully revealed. Furthermore, this unique probe reveals the sequential damage of mitochondria and lysosomes during apoptosis through the successive fading of green and red emission. Thereby, the probe enables the discrimination of health state, early apoptosis, and late apoptosis of cells with three different sets of fluorescent signals. Overall, the robust probe is a desirable molecular tool to reveal the interactions between the three organelles, and investigate cell apoptosis and relative areas.
Asunto(s)
Colorantes Fluorescentes , Orgánulos , Lisosomas , Mitocondrias , ApoptosisRESUMEN
Carboxylesterase (CEs), mainly localized in endoplasmic reticulum (ER), are responsible for hydrolyzing compounds containing various ester bonds. They have been closely associated with drug metabolism and cellular homeostasis. Although some CE fluorescent probes have been developed, there are still a lack of probes that could target to the ER. Here, we developed a novel fluorescent probe CR with a specific ER anchor for monitoring CEs. In CR, p-toluenesulfonamide was chosen for precise ER targeting. A simple acetyl moiety was used as the CE response site and fluorescence modulation unit. During the spectral tests, CR displayed a fast response speed (within 10 s) towards CEs. In addition, it showed high sensitivity [limit of detection (LOD) = 5.1 × 10-3 U/ml] and high selectivity with CEs. In biological imaging, probe CR could especially locate in the ER in HepG2 cells. After cells were treated with orilistat, CR succeeded in monitoring the changes in the CEs. Importantly, CR also had the ability to trace the changes in CEs in a tunicamycin-induced ER stress model. Therefore, probe CR could be a powerful molecular tool for further investigating the functions of CEs in the ER.
Asunto(s)
Carboxilesterasa , Colorantes Fluorescentes , Humanos , Colorantes Fluorescentes/química , Carboxilesterasa/análisis , Carboxilesterasa/química , Carboxilesterasa/metabolismo , Células HeLa , Retículo Endoplásmico/química , Retículo Endoplásmico/metabolismo , Límite de DetecciónRESUMEN
Effective and noninvasive cancer diagnosis is expected to ease the burden of continued increased deaths worldwide. Herein, we proposed viscosity of the tumor microenvironment as a biomarker and further develop a versatile optical agent, TBM-V, for monitoring tumor microenvironmental viscosity alterations to achieve cancer diagnosis, therapeutic effect tracking, and anticancer drug screening. When in highly viscous media, near-infrared signals of TBM-V are specifically activated, endowing the probe with the capacity of avoiding biological autofluorescence and achieving high signal-to-noise ratio imaging. The results of vascular imaging disclosed higher fluorescence of the blood vessels in the tumor than the normal ones, implying tumors being pointed out with brighter fluorescence. With the assistance of fluorescence imaging technology, TBM-V achieved noninvasively identifying cancer in vivo with high signal-to-noise ratio imaging. In addition, the capability of TBM-V to evaluate anticancer drug efficacy with viscosity as a robust biomarker was explored. Furthermore, as a proof of concept, screening of the anticancer drugs is also realized through in situ monitoring of the microenvironmental viscosity fluctuations of the tumor with TBM-V. Note that this proposed fluorescence imaging method outperforms the clinical hematoxylin and eosin (H&E) staining assay with the advantageous features of noninvasive and in vivo characteristics. We expected that this unique strategy will reinvigorate the continued perfection of the cancer diagnosis systems.
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Colorantes Fluorescentes/química , Neoplasias Mamarias Experimentales/diagnóstico , Espectrometría de Fluorescencia/métodos , Espectroscopía Infrarroja Corta/métodos , Viscosidad , Animales , Supervivencia Celular/efectos de los fármacos , Femenino , Colorantes Fluorescentes/toxicidad , Ratones , Ratones Endogámicos BALB C , Microambiente TumoralRESUMEN
Hydroxylamine (HA) is an important product of cell metabolism and plays a significant role in many biological processes, and therefore, real-time imaging of HA is of great importance for the in-depth study of its physiological and pathological functions. However, a HA-specific fluorescent probe is currently lacking primarily because the highly selective HA-responsive site is undeveloped. To address this critical issue, we present a HA-specific FRET-based fluorescent probe (RhChr) for the selective detection of HA in living systems. Inspired by aza-Michael addition, the unsaturated system appended with an iminium ion was employed as the new HA-specific response site. In response to HA, RhChr provided a ratiometric signal output with excellent selectivity toward HA over biothiols and ammonia. We have demonstrated that RhChr could be applied for the ratiometric imaging of endogenous HA in living cells and the evaluation of xanthine oxidase (XOD) activity in living organs.
Asunto(s)
Transferencia Resonante de Energía de Fluorescencia , Colorantes Fluorescentes , Hidroxilamina/análisis , Imagen Molecular/métodos , Imagen Óptica/métodos , Animales , Supervivencia Celular , Células Cultivadas , Humanos , Xantina Oxidasa/metabolismoRESUMEN
Emergence of fluorescence imaging with real-time and in situ manners has revolutionized the fields of tracing and defining enzymes in biological systems. ß-galactosidase is a kind of enzyme that plays vital roles in controlling multitudes of cellular functions and participating in disease pathogenesis. Thus, building fluorescent probes with high sensitivity and fidelity for visualizing ß-galactosidase in biological systems is very significative. Herein, we engineered the first ultrsensitivity ratiometric fluorescent probe CG based on ICT-FRET synergetic mechanisms for detecting ß-galactosidase. The spectrum data show that probe CG has a fast response (<20 s), as well as a very low detection limit to ß-galactosidase (0.081 U/mL). Moreover, by calculation of a serious of kinetic parameters including Km (1.42 µM), kcat (7.04 s-1), and kcat/Km (4.96 µM-1 s-1), CG demonstrates high affinity and high catalytic efficiency to ß-galactosidase. Because of its excellent water solubility, CG has well biocompatibility to visualize the ß-galactosidase in living cells. Furthermore, for imaging in bioapplications, CG is capable of detecting ß-galactosidase not only in overexpressed cell lines but also in transient expressed cell lines. Significantly, CG can monitor ß-galactosidase ex vivo selectively. We hope ongoing work to employ CG can be as an ultrasensitive powerful tool for further seeking the physiological and pathological functions in biological organisms.
Asunto(s)
Transferencia Resonante de Energía de Fluorescencia/métodos , Colorantes Fluorescentes/química , Imagen Molecular/métodos , Neoplasias Ováricas/enzimología , beta-Galactosidasa/metabolismo , Animales , Proliferación Celular , Femenino , Humanos , Límite de Detección , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Microscopía Fluorescente , Neoplasias Ováricas/patología , Células Tumorales Cultivadas , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
SO2 has been recently identified as an essential gas messenger followed by NO, CO and H2S. However, abnormal concentrations of SO2 in our bodies can cause many diseases. Thus, the real-time monitoring of SO2 to well define the generation, physiological and pathological functions of SO2 is urgently needed. In this work, we developed a novel SO2 fluorescent probe on the basis of the conjugation of semi-cyanine and coumarin derivate dyes with superior features, such as near-infrared (NIR) and two-photon dual-mode monitoring, a large Stokes shift (175 nm), ultrafast response towards SO2 (within 10 s), high selectivity and photostability. Furthermore, this probe could sense SO2 by dual colorimetric and fluorescence means. In biological imaging, the probe was able to trace exogenous and endogenous SO2 in living cells, mitochondria, E. coli, zebrafish and mice under an NIR and two-photon dual-mode. These results demonstrated that the probe has strong potential for studying the physiological and pathological functions of SO2in vitro and in vivo.
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Cumarinas/química , Colorantes Fluorescentes/química , Indoles/química , Dióxido de Azufre/análisis , Animales , Colorimetría/métodos , Cumarinas/síntesis química , Cumarinas/efectos de la radiación , Cumarinas/toxicidad , Escherichia coli , Femenino , Colorantes Fluorescentes/síntesis química , Colorantes Fluorescentes/efectos de la radiación , Colorantes Fluorescentes/toxicidad , Células Hep G2 , Humanos , Indoles/síntesis química , Indoles/efectos de la radiación , Indoles/toxicidad , Ratones Endogámicos BALB C , Microscopía Fluorescente/métodos , Imagen Óptica/métodos , Fotones , Pez CebraRESUMEN
Hydrogen peroxide (H2O2) plays important roles in the regulation of many biological processes, and the abnormal level of H2O2 has close relation with the initiation and progression of many diseases. Herein, we describe a novel red-emissive fluorescence probe (RhoB) for the visualization of H2O2 in living cells, tissues and animals. RhoB was constructed on the basis of a xanthene-based red-emissive dye, and displayed nearly no fluorescence. After the treatment with H2O2, RhoB can exhibit red fluorescence with the emission wavelength at 638 nm. RhoB exhibited highly sensitive and selective response to H2O2. Density functional theory (DFT) calculations were conducted to shed light on the optical properties of RhoB, and natural bond orbital (NBO) calculations demonstrate that the boron atom shows the highest positive electricity and further support the response mechanism. RhoB was successfully applied for imaging of exogenous and endogenous H2O2 in living cells, and also can be utilized for visualizing H2O2 in living tissues and animals.
Asunto(s)
Colorantes Fluorescentes/química , Peróxido de Hidrógeno/metabolismo , Imagen Molecular/métodos , Xantenos/química , Animales , Supervivencia Celular , Color , Colorantes Fluorescentes/metabolismo , Células HeLa , Humanos , Hígado/diagnóstico por imagen , Hígado/metabolismo , Ratones , Xantenos/metabolismoRESUMEN
Abnormal lysosomal pH is closely associated with many diseases, and real-time monitoring of lysosomal pH is important for understanding the lysosome physiological nature. Here, we present a novel lysosome-targeting two-photon fluorescent probe (MP-lys) for monitoring pH changes in living cells. As a dual site-controlled probe, MP-lys employed morpholine and piperazine groups as the lysosome-targeting groups and pH response sites. MP-lys showed rapid, reversible and sensitive fluorescence response to pH. MP-lys possessed lysosome-targeting properties, and could be used for two-photon imaging of chloroquine-induced pH variation in living cells.
Asunto(s)
Colorantes Fluorescentes/química , Lisosomas/química , Cloroquina/química , Fluorescencia , Células HeLa , Humanos , Concentración de Iones de Hidrógeno , Morfolinas/química , Fotones , Piperazina/químicaRESUMEN
1,4-Dithiothreitol (DTT) has wide applications in cell biology and biochemistry. Development of effective methods for monitoring DTT in biological systems is important for the safe handling and study of toxicity to humans. Herein, we describe a two-photon fluorescence probe (Rh-DTT) to detect DTT in living systems for the first time. Rh-DTT showed high selectivity and sensitivity to DTT. Rh-DTT can be successfully used for the two-photon imaging of DTT in living cells, and also can detect DTT in living tissues and mice.
Asunto(s)
Ditiotreitol/análisis , Colorantes Fluorescentes/química , Fotones , Xantenos/química , Animales , Supervivencia Celular , Femenino , Colorantes Fluorescentes/síntesis química , Humanos , Hígado/química , Ratones , Ratones Endogámicos BALB C , Estructura Molecular , Células Tumorales CultivadasRESUMEN
Mitochondria, as essential and interesting organelles within the eukaryotic cells, play key roles in a variety of pathologies, and its abnormalities are closely associated with Alzheimer's disease (AD) and other diseases. Studies have shown that the abnormal of viscosity and concentration of hydrogen peroxide in mitochondria were all associated with AD. Accordingly, the detection of viscosity and hydrogen peroxide in mitochondria has attracted great attention. However, it remains a great challenge to explore a single probe, which can dual-detect the viscosity and H2O2 in mitochondria. Herein, in two ways to prevent the twisted internal charge transfer (TICT) process, we designed and sythesized the first dual-detection fluorescent probe Mito-VH that can visualize viscosity and H2O2 in mitochondria with different fluorescence signals in living cells.
Asunto(s)
Colorantes Fluorescentes/química , Peróxido de Hidrógeno/metabolismo , Microscopía Fluorescente/métodos , Mitocondrias/metabolismo , Animales , Supervivencia Celular , Transporte de Electrón , Células HeLa , Humanos , Ratones , Modelos Moleculares , Conformación Molecular , Células RAW 264.7 , ViscosidadRESUMEN
Carbon monoxide (CO) is one of the most important gaseous signal molecules in biological systems. However, the investigation of the functions of CO in living organisms is restricted by the lack of functional molecular tools. To address this critical challenge, we present herein the rational design, synthesis, and inâ vivo imaging studies of a powerful two-photon excited near-infrared fluorescent probe (1-Ac) for endogenous CO monitoring. The advantageous features of the new probe include high stability, low background fluorescence, large fluorescence enhancement, high sensitivity, and two-photon excitation with emission in the near-infrared region. Significantly, these merits of the probe enable the tracking of endogenous CO in zebrafish embryos and mouse tissues for the first time.
Asunto(s)
Monóxido de Carbono/análisis , Colorantes Fluorescentes/química , Pez Cebra/metabolismo , Animales , Monóxido de Carbono/metabolismo , Embrión no Mamífero/metabolismo , Riñón/química , Riñón/metabolismo , Hígado/química , Hígado/metabolismo , Ratones , Microscopía de Fluorescencia por Excitación Multifotónica , Imagen Óptica , Células RAW 264.7 , Espectroscopía Infrarroja Corta , Pez Cebra/crecimiento & desarrolloRESUMEN
Acidic pH is a critical physiological factor for controlling the activities and functions of lysosome. Herein, we report a novel dual site-controlled and lysosome-targeted intramolecular charge transfer-photoinduced electron transfer-Fluorescence resonance energy transfer (ICT-PET-FRET) fluorescent probe (CN-pH), which was essentially the combination of a turn-on pH probe (CN-1) and a turn-off pH probe (CN-2) by a nonconjugated linker. Coumarin and naphthalimide fluorophores were selected as donor and acceptor to construct the FRET platform. Hydroxyl group and morpholine were simultaneously employed as the two pH sensing sites and controlled the fluorescence of coumarin and naphthalimide units by ICT and PET, respectively. The sensing mechanism of CN-pH to pH was essentially an integration of ICT, PET, and FRET processes. Meanwhile, the morpholine also can serve as a lysosome-targeted group. By combining the two data analysis approaches of the ratios of the two emission intensities (R) and the reverse ratio R' (R' = 1/R), the fluorescent ratio of CN-pH can show proportional relationship to pH values in a very broad range from pH 4.0 to 8.0 with high sensitivity. The probe has been successfully applied for the fluorescence imaging of the lysosomal pH values, as well as ratiometrically visualizing chloroquine-stimulated changes of intracellular pH in living cells. These features demonstrate that the probe can afford practical application in biological systems.
Asunto(s)
Transferencia Resonante de Energía de Fluorescencia , Colorantes Fluorescentes/química , Lisosomas/química , Lisosomas/metabolismo , Transporte de Electrón , Colorantes Fluorescentes/síntesis química , Células HeLa , Humanos , Concentración de Iones de Hidrógeno , Estructura Molecular , Procesos Fotoquímicos , Teoría Cuántica , Células Tumorales CultivadasRESUMEN
As one of the simplest reactive carbonyl species, formaldehyde is implicated in nervous system diseases and cancer. Organelles play crucial roles in various physiological processes in living cells. Accordingly, the detection of endogenous formaldehyde at the subcellular level is of high interest. We herein describe the development of the first organelle-targeted fluorescent formaldehyde probe (Na-FA-Lyso). The new probe exhibits favorable features including a large fluorescence enhancement (about 350-fold) and a fast response to formaldehyde. Significantly, the novel probe Na-FA-Lyso was employed to visualize the endogenous formaldehyde in the lysosomes in living cells for the first time.
Asunto(s)
Colorantes Fluorescentes/farmacología , Formaldehído/análisis , Hidrazinas/farmacología , Lisosomas/metabolismo , Naftalimidas/farmacología , Fluorescencia , Colorantes Fluorescentes/síntesis química , Colorantes Fluorescentes/efectos de la radiación , Colorantes Fluorescentes/toxicidad , Células HeLa , Humanos , Hidrazinas/síntesis química , Hidrazinas/efectos de la radiación , Hidrazinas/toxicidad , Luz , Naftalimidas/síntesis química , Naftalimidas/efectos de la radiación , Naftalimidas/toxicidadRESUMEN
Investigation of the physiological and pathological functions of formaldehyde (FA) are largely restricted by a lack of useful FA imaging agents, in particular, those that allow detection of FA in the context of living tissues. Herein, we present the rational design, synthesis, and photophysical property studies of the first two-photon fluorescent FA probe, Na-FA. Importantly, the highly desirable attributes of the probe Na-FA (such as a very large turn-on signal (up to 900-fold), a low detection limit, and a very fast onset imparted by the unique design aspects of the probe), make it possible to monitor endogenous FA in living tissues for the first time. Furthermore, sodium bisulfite was identified as a simple and convenient inhibitor of FA within biological environments.
Asunto(s)
Colorantes Fluorescentes/química , Formaldehído/metabolismo , Células HeLa , Humanos , FotonesRESUMEN
Biological uses of photosensitizers in photodynamic therapy (PDT) often suffer from a lack of tumor selectivity; a strategy based on molecule-targeted cancer therapies could provide a promising solution. To synthesize new water-soluble phthalocyanines (Pcs) for bio-conjugation with peptides or antibodies, we developed a method to synthesize asymmetrically substituted Pcs with both high water solubility and one monoamino group for conjugation with biological agents for tumor homing, using folic acid as the ligand model to direct the modified Pcs into target cells. Here, we report studies on the syntheses and characterization of these Pcs. In vitro and in vivo assays prove that the high solubility characteristic can greatly increase the tumor targeting capability of Pcs by reducing non-specific uptake. This newly designed photosensitizer accumulated almost completely in tumor regions, with a negligible signal found in other tissues in the xenograft tumor model. These initial data provide strong evidence of the high specificity tumor targeting of Pcs with folate and tri-glycerol substitutions. Theoretically, the synthesized Pcs could be conveniently conjugated to many other ligands, endorsing the broad applicability of this method for tumor-targeted PDT.
Asunto(s)
Indoles/farmacología , Neoplasias Experimentales/tratamiento farmacológico , Fotoquimioterapia , Fármacos Fotosensibilizantes/farmacología , Animales , Anticuerpos/química , Células HeLa , Humanos , Indoles/síntesis química , Indoles/química , Isoindoles , Células KB , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Estructura Molecular , Células 3T3 NIH , Péptidos/química , Fármacos Fotosensibilizantes/síntesis química , Fármacos Fotosensibilizantes/química , Solubilidad , Agua/químicaRESUMEN
A newly designed, dual-functional probe based on intracellular activation has been successfully developed for the detection of cancer cells. The probe is nearly non-fluorescent in buffer due to its highly efficient FRET quenching, but it can be specifically activated with dramatic fluorescence enhancement upon intracellular cathepsin B cleavage in target cancer cells after selective internalization via folate receptor-dependent endocytosis. Therefore, this probe enables "turn-on" visualization of cancer cells with desirable specificity and contrast enhancement. This targeted, intracellularly activatable probe exhibits low fluorescence-quenched background when compared with "always-on" probes and avoids non-specific activation by non-specifically expressed enzymes in normal tissue, which normally occurs when using common "turn on" probe design strategies. Therefore, this probe can be potentially applied in intraoperative inspection during clinical cancer surgery with higher contrast and sensitivity.
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Colorantes Fluorescentes , Espacio Intracelular/metabolismo , Imagen Molecular/métodos , Neoplasias/diagnóstico , Animales , Catepsina B , Línea Celular Tumoral , Cromatografía Líquida de Alta Presión , Citometría de Flujo , Colorantes Fluorescentes/síntesis química , Colorantes Fluorescentes/química , Ácido Fólico/metabolismo , Humanos , Ratones , Microscopía Confocal , Células 3T3 NIH , Neoplasias/patología , Plasma/metabolismo , Reproducibilidad de los Resultados , Espectrometría de Fluorescencia , Espectrofotometría Ultravioleta , Temperatura , Factores de TiempoRESUMEN
Sortase-mediated hydrazinolysis of proteins with hydrazine or its derivatives was developed for the production of recombinant protein hydrazides. This process provides an alternative approach for protein semisynthesis through the use of recombinant protein hydrazides as thioester surrogates. It also provides an alternative method for C-terminal modification of proteins with functional units as well as for the preparation of C-to-C fusion proteins.
Asunto(s)
Aminoaciltransferasas/metabolismo , Proteínas Bacterianas/metabolismo , Cisteína Endopeptidasas/metabolismo , Hidrazinas/química , Proteínas Recombinantes/metabolismo , Secuencia de Aminoácidos , Fluoresceína/química , Hidrólisis , Péptidos/química , Péptidos/metabolismo , Estructura Terciaria de Proteína , Proteínas Recombinantes/química , Anticuerpos de Dominio Único/química , Anticuerpos de Dominio Único/inmunologíaRESUMEN
Mitochondrial sulfur dioxide (SO2) plays important roles in physiological and pathological activities. Unfortunately, it is lack of a reliable tool to precisely visualize the mitochondrial SO2 and elaborate its complicated functions in various cytoactivities. Here we report a mitochondrial-immobilized fluorescent probe PM-Cl consisting of coumarin and benzyl chloride modified benzothiazole, which enables selective visualization of mitochondrial SO2via chemical immobilization. The spectral results demonstrated that probe PM-Cl could respond to SO2 with high selectivity and sensitivity. Co-localization and the fluorescence of cytolysis extraction verified the excellent mitochondrial targeting and anchoring abilities. Due to the chemical immobilization, probe PM-Cl could firmly retain into mitochondria after stimulation of carbonyl cyanide m-chlorophenyl hydrazone (CCCP) and H2O2. Significantly, a series of fluorescence images are indicative of capability for detecting the fluctuations of SO2 in mitochondria during ferroptosis. Furthermore, PM-Cl also could visualize SO2 in myocardium and muscle tissues after the stimulation of CCCP. Taken together, probe PM-Cl is a very potential molecular tool for precisely detecting mitochondrial SO2 to explore its complex functions in physiological and pathological activities.