RESUMEN
Ratiometric near-infrared fluorescent pH probes with various pKa values were innovatively designed and synthesized based on cyanine with a diamine moiety. The photochemical properties of these probes were thoroughly evaluated. Among the series, IR-PHA exhibited an optimal pKa value of approximately 6.40, closely matching the pH of cancerous tissues. This feature is particularly valuable for real-time pH monitoring in both living cells and living mice. Moreover, when administered intravenously to tumor-bearing mice, IR-PHA demonstrated rapid and significant enhancement of near-infrared fluorescence and photoacoustic signals within the tumor region. This outcome underscores the probe's exceptional capability for dual-modal cancer imaging utilizing near-infrared fluorescence (NIRF) and photoacoustic (PA) modalities. Concurrently, the application of a continuous-wave near-infrared laser efficiently ablated cancer cells in vivo, attributed to the photothermal effect induced by IR-PHA. The results strongly indicate that IR-PHA is well-suited for NIRF/PA dual-modality imaging and photothermal therapy of tumors. This makes it a promising candidate for theranostic applications involving small molecules.
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Colorantes Fluorescentes , Rayos Infrarrojos , Técnicas Fotoacústicas , Terapia Fototérmica , Animales , Técnicas Fotoacústicas/métodos , Humanos , Ratones , Colorantes Fluorescentes/química , Colorantes Fluorescentes/síntesis química , Colorantes Fluorescentes/efectos de la radiación , Terapia Fototérmica/métodos , Neoplasias/diagnóstico por imagen , Neoplasias/terapia , Concentración de Iones de Hidrógeno , Línea Celular Tumoral , Ratones Desnudos , Imagen Óptica/métodos , FemeninoRESUMEN
The use of photoswitchable fluorescent diarylethenes (fDAEs) as protein labels in fluorescence microscopy and nanoscopy has been limited by labeling inhomogeneity and the need for ultraviolet light for fluorescence activation (on-switching). To overcome these drawbacks, we prepared "turn-on mode" fDAEs featuring thienyl substituents, multiple polar residues, and a reactive maleimide group in the core structure. Conjugates with antibodies and nanobodies displayed complete on-switching and excitation with violet (405 nm) and yellow-green (<565 nm) light, respectively. Besides, they afforded high signal-to-noise ratios and low unspecific labeling in fluorescence imaging. Irradiation with visible light at 532 nm or 561 nm led to transient on-off switching ("blinking") of the fDAEs of double-labeled nanobodies so that nanoscale superresolution images were readily attained through switching and localization of individual fluorophores.
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Colorantes Fluorescentes/síntesis química , Procesos Fotoquímicos , Anticuerpos/química , Línea Celular Tumoral , Colorantes Fluorescentes/efectos de la radiación , Humanos , Maleimidas/química , Microscopía Fluorescente/métodos , Compuestos de Sulfhidrilo/química , Rayos UltravioletaRESUMEN
BACKGROUND: Zebrafish can regenerate adult cardiac tissue following injuries from ventricular apex amputation, cryoinjury, and cardiomyocyte genetic ablation. Here, we characterize cardiac regeneration from cardiomyocyte chemoptogenetic ablation caused by localized near-infrared excited photosensitizer-mediated reactive oxygen species (ROS) generation. RESULTS: Exposure of transgenic adult zebrafish, Tg(myl7:fapdl5-cerulean), to di-iodinated derivative of the cell- permeable Malachite Green ester fluorogen (MG-2I) and whole-body illumination with 660 nm light resulted in cytotoxic damage to about 30% of cardiac tissue. After chemoptogenetic cardiomyocyte ablation, heart function was compromised, and macrophage infiltration was detected, but epicardial and endocardial activation response was much muted when compared to ventricular amputation. The spared cardiomyocytes underwent proliferation and restored the heart structure and function in 45-60 days after ablation. CONCLUSIONS: This cardiomyocyte ablation system did not appear to activate the epicardium and endocardium as is noted in other cardiac injury models. This approach represents a useful model to study specifically cardiomyocyte injury, proliferation and regeneration in the absence of whole organ activation. Moreover, this system can be adapted to ablate distinct cell populations in any organ system to study their function in regeneration.
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Lesiones Cardíacas/fisiopatología , Corazón/fisiología , Regeneración/fisiología , Animales , Animales Modificados Genéticamente , Proliferación Celular/fisiología , Colorantes Fluorescentes/efectos adversos , Colorantes Fluorescentes/química , Colorantes Fluorescentes/efectos de la radiación , Corazón/efectos de los fármacos , Lesiones Cardíacas/inducido químicamente , Lesiones Cardíacas/patología , Rayos Infrarrojos/efectos adversos , Miocitos Cardíacos/patología , Colorantes de Rosanilina/efectos adversos , Colorantes de Rosanilina/química , Colorantes de Rosanilina/efectos de la radiación , Pez CebraRESUMEN
Cyanine (Cy) dyes are among the most useful organic fluorophores that have found a wide range of applications in single-molecule and super-resolution imaging as well as in other biophysical studies. However, recent observations that blueshifted derivatives of Cy dyes are formed via photoconversion have raised concerns as to the potential artifacts in multicolor imaging. Here, we report the mechanism for the photoconversion of Cy5 to Cy3 that occurs upon photoexcitation during fluorescent imaging. Our studies show that the formal C2H2 excision from Cy5 occurs mainly through an intermolecular pathway involving a combination of bond cleavage and reconstitution while unambiguously confirming the identity of the fluorescent photoproduct of Cy5 to be Cy3 using various spectroscopic tools. The carbonyl products generated from singlet oxygen-mediated photooxidation of Cy5 undergo a sequence of carbon-carbon bond-breaking and -forming events to bring about the novel dye-to-dye transformation. We also show that the deletion of a two-methine unit from the polymethine chain, which results in the formation of blueshifted products, commonly occurs in other cyanine dyes, such as Alexa Fluor 647 (AF647) and Cyanine5.5. The formation of a blueshifted congener dye can obscure the multicolor fluorescence imaging, leading to misinterpretation of the data. We demonstrate that the potentially deleterious photoconversion, however, can be exploited to develop a new photoactivation method for high-density single-particle tracking in a living cell without using UV illumination and cell-toxic additives.
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Carbocianinas/química , Colorantes Fluorescentes/química , Carbocianinas/metabolismo , Carbocianinas/efectos de la radiación , Colorantes Fluorescentes/metabolismo , Colorantes Fluorescentes/efectos de la radiación , Células HeLa , Humanos , Luz , Modelos Químicos , Oxidación-Reducción/efectos de la radiación , Procesos Fotoquímicos/efectos de la radiación , Imagen Individual de MoléculaRESUMEN
In recent years, cancer phototherapy has been extensively studied as noninvasive cancer treatment. To present efficient recognition toward cancer cells, most photosensitizers (PSs) are required to couple with tumor-targeted ligands. Interestingly, the heptamethine cyanine IR780 displays an intrinsic tumor-targeted feature even without modification. However, the photothermal efficacy and photostability of IR780 are not sufficient enough for clinical use. Herein, we involve a twisted structure of tetraphenylethene (TPE) between two molecules of IR780 to improve the photothermal conversion efficiency (PCE). The obtained molecule T780T shows strong near-infrared (NIR) fluorescence and improved PCE (38.5%) in the dispersed state. Also, the photothermal stability and ROS generation capability of T780T at the NIR range (808 nm) are both promoted. In the aqueous phase, the T780T was formulated into uniform nanoaggregates (â¼200 nm) with extremely low fluorescence and PTT response, which would reduce in vivo imaging background and side effect of PTT response in normal tissues. After intravenous injection into tumor-bearing mice, the T780T nanoaggregates display high tumor accumulation and thus remarkably inhibit the tumor growth. Moreover, the enhanced photostability of the T780T allows for twice irradiation after one injection and leads to more significant tumor inhibition. In summary, our study presents a tumor-targeted small-molecule PS for efficient cancer therapy and brings a new design of heptamethine cyanine PS for potential clinical applications.
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Antineoplásicos/uso terapéutico , Colorantes Fluorescentes/uso terapéutico , Indoles/uso terapéutico , Neoplasias/tratamiento farmacológico , Fármacos Fotosensibilizantes/uso terapéutico , Estilbenos/uso terapéutico , Animales , Antineoplásicos/síntesis química , Antineoplásicos/efectos de la radiación , Línea Celular Tumoral , Colorantes Fluorescentes/síntesis química , Colorantes Fluorescentes/efectos de la radiación , Humanos , Indoles/síntesis química , Indoles/efectos de la radiación , Rayos Infrarrojos , Ratones Endogámicos BALB C , Neoplasias/diagnóstico por imagen , Fotoquimioterapia , Fármacos Fotosensibilizantes/síntesis química , Fármacos Fotosensibilizantes/efectos de la radiación , Terapia Fototérmica , Estilbenos/síntesis química , Estilbenos/efectos de la radiaciónRESUMEN
Renal-clearable and target-responsive near-infrared (NIR) fluorescent imaging probes have been promising for in vivo diagnosis of acute kidney injury (AKI). However, designing an imaging probe that is renal-clearable and concurrently responsive toward multiple molecular targets to facilitate early detection of AKI with improved sensitivity and specificity is challenging. Herein, by leveraging the receptor-mediated binding and retention effect along with enzyme-triggered fluorescence activation, we design and synthesize an activatable small-molecule NIR fluorescent probe (1-DPA2) using a "one-pot sequential click reaction" approach. 1-DPA2 can target both the externalized phosphatidylserine (PS) and active caspase-3 (Casp-3), two essential biomarkers of apoptosis, producing enhanced 808 nm NIR fluorescence and a high signal-to-background ratio (SBR) amenable to detecting the onset of cisplatin-induced AKI in mice as early as 24 h post-treatment with cisplatin. We not only monitor the gradual activation of Casp-3 in the kidney of mice upon AKI progression but also can report on the progressive recovery of kidney functions in AKI mice following N-acetyl-l-cysteine (NAC) therapy via real-time fluorescence imaging by 1-DPA2. This study demonstrates the ability of 1-DPA2 for longitudinal monitoring of renal cell apoptosis by concurrently targeting PS externalization and Casp-3 activation, which is efficient for early diagnosis of AKI and useful for prediction of potential drug nephrotoxicity as well as in vivo screening of anti-AKI drugs' efficacy.
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Lesión Renal Aguda/diagnóstico por imagen , Caspasa 3/metabolismo , Colorantes Fluorescentes/química , Fosfatidilserinas/metabolismo , Acetilcisteína/uso terapéutico , Lesión Renal Aguda/inducido químicamente , Lesión Renal Aguda/tratamiento farmacológico , Lesión Renal Aguda/metabolismo , Animales , Apoptosis/fisiología , Biomarcadores/metabolismo , Línea Celular , Cisplatino , Complejos de Coordinación/síntesis química , Complejos de Coordinación/química , Complejos de Coordinación/efectos de la radiación , Femenino , Colorantes Fluorescentes/síntesis química , Colorantes Fluorescentes/efectos de la radiación , Indoles/síntesis química , Indoles/química , Indoles/efectos de la radiación , Rayos Infrarrojos , Ratones Endogámicos BALB C , Ratones Desnudos , Imagen Óptica , Zinc/químicaRESUMEN
Deferasirox, ExJade, is an FDA-approved iron chelator used for the treatment of iron overload. In this work, we report several fluorescent deferasirox derivatives that display unique photophysical properties, i.e., aggregation-induced emission (AIE), excited state intramolecular proton transfer, charge transfer, and through-bond and through-space conjugation characteristics in aqueous media. Functionalization of the phenol units on the deferasirox scaffold afforded the fluorescent responsive pro-chelator ExPhos, which enabled the detection of the disease-based biomarker alkaline phosphatase (ALP). The diagnostic potential of these deferasirox derivatives was supported by bacterial biofilm studies.
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Deferasirox/análogos & derivados , Colorantes Fluorescentes/química , Fosfatasa Alcalina/análisis , Antibacterianos/farmacología , Proteínas Bacterianas/análisis , Biopelículas/efectos de los fármacos , Biomarcadores/análisis , Cefoperazona/farmacología , Deferasirox/farmacología , Deferasirox/efectos de la radiación , Colorantes Fluorescentes/farmacología , Colorantes Fluorescentes/efectos de la radiación , Luz , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Staphylococcus aureus Resistente a Meticilina/enzimología , Staphylococcus aureus Resistente a Meticilina/fisiología , Pruebas de Sensibilidad Microbiana , Microscopía Confocal , Microscopía Fluorescente , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/enzimología , Pseudomonas aeruginosa/fisiología , Sulbactam/farmacologíaRESUMEN
A novel quinoline fluorescent probe QNP ((E)-N'-(5-chloro-2-hydroxybenzylidene) quinoline-2-carbohydrazide) for detection of Al3+ ion was designed, synthesized and characterized. QNP displayed a high fluorescence enhancement in the presence of Al3+ ion in DMF:PBS (99:1, v/v) solution and the detection limit was as low as 1.25 µM with high selectivity and excellent sensitivity from 0 to 3 µM. The sensing ability of QNP towards Al3+ ion is attributed to the synergistic effect of PET and ICT. Furthermore, the binding stoichiometry between QNP and Al3+ ion is of 1:1 by Job's plot and mass spectrum, and the calculated binding constant is 4.29 × 108 M-1. The detection of Al3+ ion in water samples illustrates that QNP could be applied to the detection of practical samples in the environment. Bioimaging experiments on Hela cells, zebrafish and soybean root tissues demonstrate that it has potential application to investigate biological processes involving Al3+ ion within living cells. A quinoline-based turn-on fluorescence probe for the detection of Al3+ and its bioimaging in living cells, plant, and zebrafish.
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Aluminio/análisis , Colorantes Fluorescentes/química , Hidrazonas/química , Quinolinas/química , Animales , Agua Potable/análisis , Colorantes Fluorescentes/síntesis química , Colorantes Fluorescentes/efectos de la radiación , Células HeLa , Humanos , Hidrazonas/síntesis química , Hidrazonas/efectos de la radiación , Límite de Detección , Raíces de Plantas/química , Quinolinas/síntesis química , Quinolinas/efectos de la radiación , Glycine max/química , Contaminantes Químicos del Agua/análisis , Pez CebraRESUMEN
Donor-π-acceptor (D-π-A) fluorophores consisting of a donor unit, a π linker, and an acceptor moiety have attracted attention in the last decade. In this study, we report the synthesis, characterization, optical properties, TD-DFT, and cytotoxicity studies of 17 near infrared (NIR) D-π-A analogs which have not been reported so far to the best of our knowledge. These fluorophores have chloroacrylic acid as the acceptor unit and various donor units such as indole, benzothiazole, benzo[e]indole, and quinoline. The fluorophores showed strong absorption in the NIR (700-970 nm) region due to their enhanced intramolecular charge transfer (ICT) between chloroacrylic acid and the donor moieties connected with the Vilsmeier-Haack linker. The emission wavelength maxima of the fluorophores were in between 798 and 870 nm. Compound 20 with a 4-quinoline donor moiety showed an emission wavelength above 1000 nm in the NIR II window. The synthesized fluorophores were characterized by 1H NMR and 13C NMR, and their optical properties were studied. Time dependent density functional theory (TD-DFT) calculations showed that the charge transfer occurs from the donor groups (indole, benzothiazole, benzo[e]indole, and quinoline) to the acceptor chloroacrylic acid moiety. Fluorophores with [HOMO] to [LUMO+1] transitions were shown to possess a charge separation character. The cytotoxicity of selected fluorophores, 4, 7, 10 and 12 was investigated against breast cancer cell lines and they showed better activity than the anti-cancer agent docetaxel.
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Acrilatos/farmacología , Antineoplásicos/farmacología , Colorantes Fluorescentes/farmacología , Compuestos Heterocíclicos con 2 Anillos/farmacología , Acrilatos/síntesis química , Acrilatos/efectos de la radiación , Antineoplásicos/síntesis química , Antineoplásicos/efectos de la radiación , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Teoría Funcional de la Densidad , Ensayos de Selección de Medicamentos Antitumorales , Estabilidad de Medicamentos , Colorantes Fluorescentes/síntesis química , Colorantes Fluorescentes/efectos de la radiación , Compuestos Heterocíclicos con 2 Anillos/síntesis química , Compuestos Heterocíclicos con 2 Anillos/efectos de la radiación , Humanos , Luz , Modelos Químicos , Fenómenos ÓpticosRESUMEN
Super-resolution fluorescence microscopy has become an important catalyst for discovery in the life sciences. In STimulated Emission Depletion (STED) microscopy, a pattern of light drives fluorophores from a signal-emitting on-state to a non-signalling off-state. Only emitters residing in a sub-diffraction volume around an intensity minimum are allowed to fluoresce, rendering them distinguishable from the nearby, but dark fluorophores. STED routinely achieves resolution in the few tens of nanometers range in biological samples and is suitable for live imaging. Here, we review the working principle of STED and provide general guidelines for successful STED imaging. The strive for ever higher resolution comes at the cost of increased light burden. We discuss techniques to reduce light exposure and mitigate its detrimental effects on the specimen. These include specialized illumination strategies as well as protecting fluorophores from photobleaching mediated by high-intensity STED light. This opens up the prospect of volumetric imaging in living cells and tissues with diffraction-unlimited resolution in all three spatial dimensions.
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Procesamiento de Imagen Asistido por Computador/métodos , Microscopía Fluorescente/métodos , Color , Equipo Reutilizado , Fluorescencia , Colorantes Fluorescentes/química , Colorantes Fluorescentes/efectos de la radiación , Iluminación/métodos , Imagen Óptica/métodos , Fotoblanqueo , Error Científico Experimental , Factores de TiempoRESUMEN
A visible-light-mediated, mild and one-pot three-component reaction in the presence of organophotoredox catalyst Eosin Y using EtOH:H2O as reaction medium for the synthesis of 3-functionalized indole derivatives was developed. Visible light used in the protocol is green, inexpensive, readily available energy source. The sustainable reagents make the protocol compatible with green chemistry demands.
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Eosina Amarillenta-(YS)/efectos de la radiación , Colorantes Fluorescentes/efectos de la radiación , Indoles/síntesis química , Luz , Catálisis , Eosina Amarillenta-(YS)/química , Colorantes Fluorescentes/químicaRESUMEN
The Ti3C2 MXene quantum dots (Ti3C2 MQDs) derived from Ti3C2 MXene have received much attention because of their remarkable advantages in biosensing. Nevertheless, the functionalization of Ti3C2 MQDs to improve their properties is just in its infant stage. Herein, we firstly synthesized nitrogen and boron co-doped Ti3C2 MQDs (N, B-Ti3C2 MQDs) with good water solubility, strong stability, and high optical characteristics. The N, B-Ti3C2 MQDs exhibit excitation wavelength-dependent blue photoluminescence with optimal excitation/emission peaks at 335/439 nm. Nowadays, the development of fast and real-time detection of tetracycline (TC) in animal derived food is very essential. In this work, a novel point-of-care testing (POCT) platform was established based on ratiometric fluorescence method using N, B-Ti3C2 MQDs coupled with Eu3+. Upon addition of TC in the Eu3+/N, B-MQDs system, blue fluorescence emission of N, B-Ti3C2 MQDs was quenched and red fluorescence emission of Eu3+ was enhanced gradually, which was ascribed to the synergistic inner filter effect and antenna effect. Moreover, we prepared test papers with N, B-Ti3C2 MQDs and Eu3+ for TC detection based on the change of fluorescence color, which could be recognized by color recognizer app installed in the smartphone. Therefore, great promise for POCT of TC is given with the merits of simplicity and visible detection possibility. The proposed method demonstrated a low detection limit of 20 nM. Application of the platform for TC quantification in milk samples opened a novel means for the potential use of N, B-Ti3C2 MQDs in food safety.
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Europio/química , Colorantes Fluorescentes/química , Pruebas en el Punto de Atención , Puntos Cuánticos/química , Tetraciclina/análisis , Titanio/química , Animales , Antibacterianos/análisis , Boro/química , Boro/efectos de la radiación , Europio/efectos de la radiación , Fluorescencia , Colorantes Fluorescentes/efectos de la radiación , Contaminación de Alimentos/análisis , Límite de Detección , Leche/química , Nitrógeno/química , Nitrógeno/efectos de la radiación , Papel , Puntos Cuánticos/efectos de la radiación , Teléfono Inteligente , Espectrometría de Fluorescencia/instrumentación , Espectrometría de Fluorescencia/métodos , Titanio/efectos de la radiación , Rayos UltravioletaRESUMEN
Near-infrared (NIR) activatable fluorescent probes have been considered to be the effective edge tools for the investigation of cell biology and disease diagnosis because of their outstanding advantages. Related genes involved in tumor genesis and progression regulate the overexpression of certain enzymes. Owing to the distinctive characteristics of quick reaction time and favorable pharmacokinetics, enzyme-reactive NIR optical probes have shown great potential in the diagnosis of tumorigenesis and in image-guided intraoperative surgeries with high signal-to-noise ratios. In this review, we mainly summarize the latest advancements in enzyme-reactive NIR fluorescent probes from design strategy to biomedical application. Specifically, some challenges and prospects in this field are presented at the end of the review, which will be beneficial to innovatively construct new multifunctional fluorescent probes and actively promote their clinical transformation in the future.
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Enzimas/metabolismo , Colorantes Fluorescentes/química , Neoplasias/diagnóstico por imagen , Animales , Antineoplásicos/química , Antineoplásicos/metabolismo , Antineoplásicos/efectos de la radiación , Antineoplásicos/uso terapéutico , Línea Celular Tumoral , Enzimas/química , Colorantes Fluorescentes/metabolismo , Colorantes Fluorescentes/efectos de la radiación , Colorantes Fluorescentes/uso terapéutico , Humanos , Rayos Infrarrojos , Estructura Molecular , Neoplasias/tratamiento farmacológico , Neoplasias/enzimología , Cirugía Asistida por Computador/métodosRESUMEN
Biodegradable nanostructures displaying aggregation-induced emission (AIE) are desirable from a biomedical point of view, due to the advantageous features of loading capacity, emission brightness, and fluorescence stability. Herein, biodegradable polymers comprising poly (ethylene glycol)-block-poly(caprolactone-gradient-trimethylene carbonate) (PEG-P(CLgTMC)), with tetraphenylethylene pyridinium-TMC (PAIE) side chains have been developed, which self-assembled into well-defined polymersomes. The resultant AIEgenic polymersomes are intrinsically fluorescent delivery vehicles. The presence of the pyridinium moiety endows the polymersomes with mitochondrial targeting ability, which improves the efficiency of co-encapsulated photosensitizers and improves therapeutic index against cancer cells both in vitro and in vivo. This contribution showcases the ability to engineer AIEgenic polymersomes with structure inherent fluorescence and targeting capacity for enhanced photodynamic therapy.
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Antineoplásicos/farmacología , Plásticos Biodegradables/farmacología , Colorantes Fluorescentes/farmacología , Fármacos Fotosensibilizantes/farmacología , Poliésteres/farmacología , Polietilenglicoles/farmacología , Antineoplásicos/síntesis química , Antineoplásicos/efectos de la radiación , Compuestos de Bencilideno/síntesis química , Compuestos de Bencilideno/farmacología , Compuestos de Bencilideno/efectos de la radiación , Plásticos Biodegradables/síntesis química , Plásticos Biodegradables/efectos de la radiación , Compuestos de Boro/síntesis química , Compuestos de Boro/farmacología , Compuestos de Boro/efectos de la radiación , Línea Celular Tumoral , Colorantes Fluorescentes/síntesis química , Colorantes Fluorescentes/efectos de la radiación , Humanos , Luz , Fármacos Fotosensibilizantes/síntesis química , Fármacos Fotosensibilizantes/efectos de la radiación , Poliésteres/síntesis química , Poliésteres/efectos de la radiación , Polietilenglicoles/síntesis química , Polietilenglicoles/efectos de la radiación , Compuestos de Piridinio/síntesis química , Compuestos de Piridinio/farmacología , Compuestos de Piridinio/efectos de la radiaciónRESUMEN
Focused ultrasound has been shown to be effective at stimulating neurons in many animal models, both in vivo and ex vivo Ultrasonic neuromodulation is the only noninvasive method of stimulation that could reach deep in the brain with high spatial-temporal resolution, and thus has potential for use in clinical applications and basic studies of the nervous system. Understanding the physical mechanism by which energy in a high acoustic frequency wave is delivered to stimulate neurons will be important to optimize this technology. We imaged the isolated salamander retina of either sex during ultrasonic stimuli that drive ganglion cell activity and observed micron scale displacements, consistent with radiation force, the nonlinear delivery of momentum by a propagating wave. We recorded ganglion cell spiking activity and changed the acoustic carrier frequency across a broad range (0.5-43 MHz), finding that increased stimulation occurs at higher acoustic frequencies, ruling out cavitation as an alternative possible mechanism. A quantitative radiation force model can explain retinal responses and could potentially explain previous in vivo results in the mouse, suggesting a new hypothesis to be tested in vivo Finally, we found that neural activity was strongly modulated by the distance between the transducer and the electrode array showing the influence of standing waves on the response. We conclude that radiation force is the dominant physical mechanism underlying ultrasonic neurostimulation in the ex vivo retina and propose that the control of standing waves is a new potential method to modulate these effects.SIGNIFICANCE STATEMENT Ultrasonic neurostimulation is a promising noninvasive technology that has potential for both basic research and clinical applications. The mechanisms of ultrasonic neurostimulation are unknown, making it difficult to optimize in any given application. We studied the physical mechanism by which ultrasound is converted into an effective energy form to cause neurostimulation in the retina and find that ultrasound acts via radiation force leading to a mechanical displacement of tissue. We further show that standing waves have a strong modulatory effect on activity. Our quantitative model by which ultrasound generates radiation force and leads to neural activity will be important in optimizing ultrasonic neurostimulation across a wide range of applications.
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Retina/efectos de la radiación , Ondas Ultrasónicas , Acústica , Potenciales de Acción/efectos de la radiación , Ambystoma , Animales , Femenino , Colorantes Fluorescentes/efectos de la radiación , Masculino , Ratones , Microscopía Confocal , Modelos Neurológicos , Técnicas de Cultivo de Órganos , Fosfenos/fisiología , Compuestos de Piridinio/efectos de la radiación , Compuestos de Amonio Cuaternario/efectos de la radiación , Células Ganglionares de la Retina/fisiología , Células Ganglionares de la Retina/efectos de la radiación , TemperaturaRESUMEN
Photophysics tunability through alteration of framework aperture (metal-organic framework (MOF) = variable; guest = constant) was probed for the first time in comparison with previously explored concepts (MOF = constant; guest = variable). In particular, analysis of the confinement effect on a photophysical response of integrated 5-(3-chlorobenzylidene)-2,3-dimethyl-3,5-dihydro-4H-imidazol-4-one (Cl-BI) chromophore allowed us to establish a photophysics-aperture relationship. To shed light on the observed correlation, the framework confined environment was replicated using a molecular cage, Pd6(TPT)4 (TPT = 2,4,6-tri(pyridin-4-yl)-1,3,5-triazine), thus allowing for utilization of crystallography, spectroscopy, and theoretical simulations to reveal the effect a confined space has on the chromophore's molecular conformation (including disruption of strong hydrogen bonding and novel conformer formation) and any associated changes on a photophysical response. Furthermore, the chosen Cl-oHBI@Pd6(TPT)4 (Cl-oHBI = 5-(5-chloro-2-hydroxybenzylidene)-2,3-dimethyl-3,5-dihydro-4H-imidazol-4-one, chromophore) system was applied as a tool for targeted cargo delivery of a chromophore to the confined space of DNA, which resulted in promotion of chromophore-DNA interactions through a well-established intercalation mechanism. Moreover, the developed principles were applied toward utilizing a HBI-based chromophore as a fluorescent probe on the example of macrophage cells. For the first time, suppression of non-radiative decay pathways of a chromophore was tested by anchoring the chromophore to a framework metal node, portending a potential avenue to develop an alternative to natural biomarkers. Overall, these studies are among the first attempts to demonstrate the unrevealed potential of a confined scaffold environment for tailoring a material's photophysical response.
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ADN/química , Colorantes Fluorescentes/química , Imidazoles/química , Estructuras Metalorgánicas/química , Triazinas/química , Colorantes Fluorescentes/efectos de la radiación , Células HeLa , Humanos , Imidazoles/efectos de la radiación , Sustancias Intercalantes/química , Sustancias Intercalantes/efectos de la radiación , Luz , Conformación MolecularRESUMEN
Photoactivation of bioactive molecules allows manipulation of cellular processes with high spatiotemporal precision. The recent emergence of visible-light excitable photoprotecting groups has the potential to further expand the established utility of the photoactivation strategy in biological applications by offering higher tissue penetration, diminished phototoxicity, and compatibility with other light-dependent techniques. Nevertheless, a critical barrier to such applications remains the significant hydrophobicity of most visible-light excitable photocaging groups. Here, we find that applying the conventional 2,6-sulfonation to meso-methyl BODIPY photocages is incompatible with their photoreaction due to an increase in the excited state barrier for photorelease. We present a simple, remote sulfonation solution to BODIPY photocages that imparts water solubility and provides control over cellular permeability while retaining their favorable spectroscopic and photoreaction properties. Peripherally disulfonated BODIPY photocages are cell impermeable, making them useful for modulation of cell-surface receptors, while monosulfonated BODIPY retains the ability to cross the cellular membrane and can modulate intracellular targets. This new approach is generalizable for controlling BODIPY localization and was validated by sensitization of mammalian cells and neurons by visible-light photoactivation of signaling molecules.
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Alcanosulfonatos/metabolismo , Compuestos de Boro/metabolismo , Colorantes Fluorescentes/metabolismo , Alcanosulfonatos/síntesis química , Alcanosulfonatos/efectos de la radiación , Animales , Compuestos de Boro/síntesis química , Compuestos de Boro/efectos de la radiación , Membrana Celular/metabolismo , Dopamina/química , Dopamina/farmacología , Portadores de Fármacos/síntesis química , Portadores de Fármacos/metabolismo , Portadores de Fármacos/efectos de la radiación , Colorantes Fluorescentes/síntesis química , Colorantes Fluorescentes/efectos de la radiación , Células HEK293 , Hipocampo/efectos de los fármacos , Histamina/química , Histamina/farmacología , Humanos , Luz , Microscopía Confocal , Microscopía Fluorescente , Estructura Molecular , Neuronas/efectos de los fármacos , Ratas , SolubilidadRESUMEN
Detecting myeloperoxidase (MPO) activity in living organisms is important because MPO contributes to the pathogenesis of many diseases such as rheumatoid arthritis and other inflammatory diseases, artherosclerosis, neurodegenerative disease, and some cancers. However, rapid and effective methods for the detection of basal MPO activity in living systems have not yet been reported. Herein, we report a near-infrared (NIR) emissive "turn-on" probe FD-301 that can specifically bind to MPO and accurately measure MPO activity in living cells and in vivo via a rapid response to initial hypochlorous acid (HOCl), produced by MPO. Notably, FD-301 could detect the basal level of MPO activity in human promyelocytic leukemia cells (HL-60) and could discriminate between MPO high-expression and low-expression cells. Furthermore, FD-301 was successfully applied to in vivo imaging of MPO in MPO-dependent diseases, such as arthritis and inflammatory bowel disease.
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Colorantes Fluorescentes/química , Peroxidasa/análisis , Fenotiazinas/química , Enfermedad Aguda , Animales , Artritis/enzimología , Colon/patología , Colorantes Fluorescentes/metabolismo , Colorantes Fluorescentes/efectos de la radiación , Células HL-60 , Humanos , Ácido Hipocloroso/metabolismo , Enfermedades Inflamatorias del Intestino/enzimología , Enfermedades Inflamatorias del Intestino/patología , Rayos Infrarrojos , Masculino , Ratones , Imagen Óptica , Peroxidasa/metabolismo , Fenotiazinas/metabolismo , Fenotiazinas/efectos de la radiación , Unión Proteica , Células RAW 264.7RESUMEN
Lipid droplets (LDs) are organelles that play a major role in regulating the storage of neutral lipids. Dysregulation of LDs is associated with metabolic disorders, such as fatty liver diseases, obesity, diabetes, and atherosclerosis. We have developed LD-selective small-molecule fluorescence probes (probes 3 and 4) that are available for both one- and two-photon microscopy, employing live or fixed cells. We found that probes 3 and 4 sensitively detect the increased LDs in response to oleic acid or endoplasmic reticulum stress, both in cells and tissues of the liver. The narrow absorption and emission bands of probes 3 and 4 allow multicolor imaging for the study of the role of LDs in pathophysiology and LD-associated signaling by the coapplication of the probes for different organelles or antibodies against specific proteins. In addition, we show here, for the first time, that two-photon microscopy imaging using our LD-selective probes with LysoTracker provides a novel method for screening drugs to potentially induce steatosis and/or phospholipidosis.
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Hígado Graso/diagnóstico por imagen , Colorantes Fluorescentes/química , Gotas Lipídicas/metabolismo , Lipidosis/diagnóstico por imagen , Animales , Benzofuranos/síntesis química , Benzofuranos/química , Benzofuranos/efectos de la radiación , Estrés del Retículo Endoplásmico/efectos de los fármacos , Hígado Graso/inducido químicamente , Colorantes Fluorescentes/síntesis química , Colorantes Fluorescentes/efectos de la radiación , Células HeLa , Humanos , Lipidosis/inducido químicamente , Ratones , Microscopía Fluorescente , FotonesRESUMEN
Ferroptosis is an iron-dependent form of regulated cell death. In this study, a ratiometric fluorescent probe, gold carbon dots (GCDs) consisting of carbon skeleton and gold nanoclusters, was used for in situ imaging to monitor redox status in biothiols (glutathione and cysteine) and ferric metabolism of cancer cells in ferroptosis. The as-prepared GCDs can selectively respond to biothiols, interestingly, the fluorescence may be switched to sense ferric ions without interference by biothiols under proper conditions. The robust GCDs-probe exhibits excellent photobleaching resistance and can reversibly respond to intracellular biothiols/ferric ion with high temporal resolution. The 8 h real-time imaging of living cells was employed to track the fluctuation of biothiols, showing the change of redox status in ferroptosis. In addition, release of ferric ions in cells was monitored. The real-time imaging of depletion of biothiols and release of ferric ion in cells indicates the GCDs-probe can monitor how the ferroptosis regulates redox status in biothiols and ferric metabolism.