Specific two-photon imaging of live cellular and deep-tissue lipid droplets by lipophilic AIEgens at ultra-low concentration

Lipid droplets are highly associated with obesity, diabetes, inflammatory disorders and cancer. A reliable two-photon dye for specific lipid droplets imaging in live cells and live tissues at ultra-low concentration has rarely been reported. In this work, four new aggregation-induced emission luminogens (AIEgens) based on the naphthalene core were designed and synthesized for specific two-photon lipid droplets staining. The new molecules, namely NAP AIEgens, exhibit large Stokes shift (>110 nm), high solid-state fluorescence quantum yield (up to 30%), good two-photon absorption cross section (45–100 GM at 860 nm), high biocompatibility and good photostability. They could specifically stain lipid droplets at ultra-low concentration (50 nM) in a short time of 15 min. Such ultra-low concentration is the lowest value for lipid droplets staining in live cells reported so far. In vitro and ex vivo two-photon imaging of lipid droplets in live cells and live mice liver tissues were successfully demonstrated. In addition, selective visualization of lipid droplets in live mice liver tissues could be achieved at a depth of about 70 μm. These excellent properties render them as promising candidates for investigating lipid droplets-associated physiological and pathological processes in live biological samples

Cancer-cell-specific Self-Reporting Photosensitizer for Precise Identification and Ablation of Cancer Cells

Cancer-cell-specific fluorescent photosensitizers (PSs) are highly desired molecular tools for cancer ablation with minimal damage to normal cells. However, such PSs that can achieve cancer specification and ablation and a self-reporting manner concurrently are rarely reported and still an extremely challenging task. Herein, we have proposed a feasible strategy and conceived a series of fluorescent PSs based on simple chemical structures for identifying and killing cancer cells as well as monitoring the photodynamic therapy (PDT) process by visualizing the change of subcellular localization. All of the constructed cationic molecules could stain mitochondria in cancer cells, identify cancer cells specifically, and monitor cancer cell viability. Among these, IVP-Br has the strongest ability to produce ROS, which serves as a potent PS for specific recognition and killing of cancer cells. IVP-Br could translocate from mitochondria to the nucleolus during PDT, self-reporting the entire therapeutic process. Mechanism study confirms that IVP-Br with light irradiation causes cancer cell ablation via inducing cell cycle arrest, cell apoptosis, and autophagy. The efficient ablation of tumor through PDT induced by IVP-Br has been confirmed in the 3D tumor spheroid chip. Particularly, IVP-Br could discriminate cancer cells from white blood cells (WBCs), exhibiting great potential to identify circulating tumor cells (CTCs)

Cancer-cell-specific Self-Reporting Photosensitizer for Precise Identification and Ablation of Cancer Cells

Cancer-cell-specific fluorescent photosensitizers (PSs) are highly desired molecular tools for cancer ablation with minimal damage to normal cells. However, such PSs that can achieve cancer specification and ablation and a self-reporting manner concurrently are rarely reported and still an extremely challenging task. Herein, we have proposed a feasible strategy and conceived a series of fluorescent PSs based on simple chemical structures for identifying and killing cancer cells as well as monitoring the photodynamic therapy (PDT) process by visualizing the change of subcellular localization. All of the constructed cationic molecules could stain mitochondria in cancer cells, identify cancer cells specifically, and monitor cancer cell viability. Among these, IVP-Br has the strongest ability to produce ROS, which serves as a potent PS for specific recognition and killing of cancer cells. IVP-Br could translocate from mitochondria to the nucleolus during PDT, self-reporting the entire therapeutic process. Mechanism study confirms that IVP-Br with light irradiation causes cancer cell ablation via inducing cell cycle arrest, cell apoptosis, and autophagy. The efficient ablation of tumor through PDT induced by IVP-Br has been confirmed in the 3D tumor spheroid chip. Particularly, IVP-Br could discriminate cancer cells from white blood cells (WBCs), exhibiting great potential to identify circulating tumor cells (CTCs)

Spatially Dependent Fluorescent Probe for Detecting Different Situations of Mitochondrial Membrane Potential Conveniently and Efficiently

The feedback from mitochondrial membrane potential (MMP) in different situations (normal, decreasing, and vanishing) can reflect different cellular status, which can be applied in biomedical research and diagnosis of the related diseases. Thus, the efficient and convenient detection for MMP in different situations is particularly important, yet the operations of current fluorescent probes are complex. In order to address this concern, we presented herein a spatially dependent fluorescent probe composed of organic cationic salt. The experimental results from normal and immortalized cells showed that it could accumulate in mitochondria selectively when MMP was normal. Also, it would move into the nucleus from mitochondria gradually with the decrease of MMP, and finally it targeted the nucleus exclusively when MMP vanished. According to the cell morphology, there is a straightforward spatial boundary between the nucleus and cytoplasm where mitochondria locate; thus, the three situations of MMP can be point-to-point indicated just by fluorescence images of the probe: that all probes accumulate in mitochondria corresponds to normal MMP; that probes locate both in the mitochondria and nucleus corresponds to decreasing MMP; that probes only target the nucleus corresponds to vanishing MMP. It is worth noting that counterstaining results with S-11348 indicated that the spatially dependent probe could be applied to distinguishing dead from viable cells in the same cell population. Compared with the commercial Cellstain-Double staining kit containing calcein-AM and propidium iodide (PI), this probe can address this concern by itself and shorten the testing time, which brings enormous convenience for relevant researches

Spatially Dependent Fluorescent Probe for Detecting Different Situations of Mitochondrial Membrane Potential Conveniently and Efficiently

The feedback from mitochondrial membrane potential (MMP) in different situations (normal, decreasing, and vanishing) can reflect different cellular status, which can be applied in biomedical research and diagnosis of the related diseases. Thus, the efficient and convenient detection for MMP in different situations is particularly important, yet the operations of current fluorescent probes are complex. In order to address this concern, we presented herein a spatially dependent fluorescent probe composed of organic cationic salt. The experimental results from normal and immortalized cells showed that it could accumulate in mitochondria selectively when MMP was normal. Also, it would move into the nucleus from mitochondria gradually with the decrease of MMP, and finally it targeted the nucleus exclusively when MMP vanished. According to the cell morphology, there is a straightforward spatial boundary between the nucleus and cytoplasm where mitochondria locate; thus, the three situations of MMP can be point-to-point indicated just by fluorescence images of the probe: that all probes accumulate in mitochondria corresponds to normal MMP; that probes locate both in the mitochondria and nucleus corresponds to decreasing MMP; that probes only target the nucleus corresponds to vanishing MMP. It is worth noting that counterstaining results with S-11348 indicated that the spatially dependent probe could be applied to distinguishing dead from viable cells in the same cell population. Compared with the commercial Cellstain-Double staining kit containing calcein-AM and propidium iodide (PI), this probe can address this concern by itself and shorten the testing time, which brings enormous convenience for relevant researches

El Volapük : revista mensual literaria y científica: Número I - 1888 enero 1

Copia digital. Madrid : Ministerio de Cultura. Subdirección General de Coordinación Bibliotecaria, 200

Interface-Targeting Strategy Enables Two-Photon Fluorescent Lipid Droplet Probes for High-Fidelity Imaging of Turbid Tissues and Detecting Fatty Liver

Lipid droplets (LDs) with unique interfacial architecture not only play crucial roles in protecting a cell from lipotoxicity and lipoapoptosis but also closely relate with many diseases such as fatty liver and diabetes. Thus, as one of the important applied biomaterials, fluorescent probes with ultrahigh selectivity for in situ and high-fidelity imaging of LDs in living cells and tissues are critical to elucidate relevant physiological and pathological events as well as detect related diseases. However, available probes only utilizing LDs’ waterless neutral cores but ignoring the unique phospholipid monolayer interfaces exhibit low selectivity. They cannot differentiate neutral cores of LDs from intracellular other lipophilic microenvironments, which results in extensively cloud-like background noise and severely limited their bioapplications. Herein, to design LD probes with ultrahigh selectivity, the exceptional interfacial architecture of LDs is considered adequately and thus an interface-targeting strategy is proposed for the first time. According to the novel strategy, we have developed two amphipathic fluorescent probes (<b>N-Cy</b> and <b>N-Py</b>) by introducing different cations into a lipophilic fluorophore (nitrobenzoxadiazole (NBD)). Consequently, their cationic moiety precisely locates the interfaces through electrostatic interaction and simultaneously NBD entirely embeds into the waterless core via hydrophobic interaction. Thus, high-fidelity and background-free fluorescence imaging of LDs are expectably realized in living cells in situ. Moreover, LDs in turbid tissues like skeletal muscle slices have been clearly imaged (up to 82 μm depth) by a two-photon microscope. Importantly, using <b>N-Cy</b>, we not only intuitively monitored the variations of LDs in number, size, and morphology but also clearly revealed their abnormity in hepatic tissues resulting from fatty liver. Therefore, these unique probes provide excellent imaging tools for elucidating LD-related physiological and pathological processes and the interface-targeting strategy possesses universal significance for designing probes with ultrahigh selectivity

Phospholipid-Biomimetic Fluorescent Mitochondrial Probe with Ultrahigh Selectivity Enables In Situ and High-Fidelity Tissue Imaging

In situ and directly imaging mitochondria in tissues instead of isolated cells can offer more native and accurate information. Particularly, in the clinical diagnose of mitochondrial diseases such as mitochondrial myopathy, it is a routine examination item to directly observe mitochondrial morphology and number in muscle tissues from patients. However, it is still a challenging task because the selectivity of available probes is inadequate for exclusively tissue imaging. Inspired by the chemical structure of amphiphilic phospholipids in mitochondrial inner membrane, we synthesized a phospholipid-biomimetic amphiphilic fluorescent probe (<b>Mito-MOI</b>) by modifying a C₁₈-alkyl chain to the lipophilic side of carbazole-indolenine cation. Thus, the phospholipid-like <b>Mito-MOI</b> locates at mitochondrial inner membrane through electrostatic interaction between its cation and inner membrane negative charge. Simultaneously, the C₁₈-alkyl chain, as the second targeting group, is deeply embedded into the hydrophobic region of inner membrane through hydrophobic interaction. Therefore, the dual targeting groups (cation and C₁₈-alkyl chain) actually endow <b>Mito-MOI</b> with ultrahigh selectivity. As expected, high-resolution microscopic photos showed that <b>Mito-MOI</b> indeed stained mitochondrial inner membrane. Moreover, in situ and high-fidelity tissue imaging has been achieved, and particularly, four kinds of mitochondria and their crystal-like structure in muscle tissues were visualized clearly. Finally, the dynamic process of mitochondrial fission in living cells has been shown. The strategy employing dual targeting groups should have reference value for designing fluorescent probes with ultrahigh selectivity to various intracellular membranous components

Specific Two-Photon Imaging of Live Cellular and Deep-Tissue Lipid Droplets by Lipophilic AIEgens at Ultralow Concentration

Lipid droplets are highly associated with obesity, diabetes, inflammatory disorders, and cancer. A reliable two-photon dye for specific lipid droplets imaging in live cells and live tissues at ultralow concentration has rarely been reported. In this work, four new aggregation-induced emission luminogens (AIEgens) based on the naphthalene core were designed and synthesized for specific two-photon lipid droplet staining. The new molecules, namely, NAP AIEgens, exhibit large Stokes shift (>110 nm), high solid-state fluorescence quantum yield (up to 30%), good two-photon absorption cross section (45–100 GM at 860 nm), high biocompatibility, and good photostability. They could specifically stain lipid droplets at ultralow concentration (50 nM) in a short time of 15 min. Such ultralow concentration is the lowest value for lipid droplets staining in live cells reported so far. <i>In vitro</i> and <i>ex vivo</i> two-photon imaging of lipid droplets in live cells and live mice liver tissues were successfully demonstrated. In addition, selective visualization of lipid droplets in live mice liver tissues could be achieved at a depth of about 70 μm. These excellent properties render them as promising candidates for investigating lipid droplet-associated physiological and pathological processes in live biological samples

Specific Two-Photon Imaging of Live Cellular and Deep-Tissue Lipid Droplets by Lipophilic AIEgens at Ultralow Concentration

Lipid droplets are highly associated with obesity, diabetes, inflammatory disorders, and cancer. A reliable two-photon dye for specific lipid droplets imaging in live cells and live tissues at ultralow concentration has rarely been reported. In this work, four new aggregation-induced emission luminogens (AIEgens) based on the naphthalene core were designed and synthesized for specific two-photon lipid droplet staining. The new molecules, namely, NAP AIEgens, exhibit large Stokes shift (>110 nm), high solid-state fluorescence quantum yield (up to 30%), good two-photon absorption cross section (45–100 GM at 860 nm), high biocompatibility, and good photostability. They could specifically stain lipid droplets at ultralow concentration (50 nM) in a short time of 15 min. Such ultralow concentration is the lowest value for lipid droplets staining in live cells reported so far. <i>In vitro</i> and <i>ex vivo</i> two-photon imaging of lipid droplets in live cells and live mice liver tissues were successfully demonstrated. In addition, selective visualization of lipid droplets in live mice liver tissues could be achieved at a depth of about 70 μm. These excellent properties render them as promising candidates for investigating lipid droplet-associated physiological and pathological processes in live biological samples