- 标题
- 摘要
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- 实验方案
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Peptic Fluorescent “Signal-On” and “Signal-Off” Sensors Utilized for the Detection Protein Post-Translational Modifications
摘要: Protein post-translational modifications (PTMs) are typically enzyme-catalyzed events generating functional diversi?cation of proteome; thus, multiple PTM enzymes have been validated as potential drug targets. We have previously introduced energy-transfer-based signal-modulation method called quenching resonance energy transfer (QRET), and utilize it to monitor PTM addition or removal using the developed peptide-break technology. Now we have reinvented the QRET technology, and as a model, we introduced the tunable ?uorescent “signal-on” and “signal-o?” detection scheme in the peptide-break PTM detection. Taking the advantage of time-resolved ?uorescence-based single-label detection technology, we were able to select the signal direction upon PTM addition or removal by simply introducing di?erent soluble Eu3+-signal-modulating molecule. This enables the selection of positive signal change upon measurable event, without any additional labeling steps, changes in assay condition or Eu3+-reporter. The concept functionality was demonstrated with four Eu3+-signal modulators in a high-throughput compatible kinase and phosphatase assays using signal-on and signal-o? readout at 615 nm or time-resolved Fo?rster resonance energy transfer at 665 nm. Our data suggest that the introduced signal modulation methodology provides a transitional ?uorescence-based single-label detection concept not limited only to PTM detection.
关键词: time-resolved fluorescence,signal-off,high-throughput screening,peptide-break technology,protein post-translational modifications,FRET,signal-on
更新于2025-11-19 16:56:35
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Black phosphorous sensitized TiO <sub/>2</sub> mesocrystals photocatalyst for hydrogen evolution with visible and near-infrared light irradiation
摘要: Wide absorption from ultraviolet (UV) to near-infrared (NIR) region and enhanced charge separation are two main requirements for promising semiconductor photocatalysts. Here, we studied visible-NIR driven photocatalytic hydrogen evolution over black phosphorus nanosheets/TiO2 mesocrystals loaded with Pt heterostructure (BP NS/ Pt (3 wt%)/TMC TMC). BP NS/Pt (3 wt%)/TMC can harvest photons from UV to NIR and simultaneously has enhanced charge separation to increase the generation of electrons for photocatalytic reduction of water. BP NS/Pt (3 wt%)/TMC exhibited photocatalytic H2 evolution rates of 1.9 and 0.41 μmol h-1 under visible (λ> 420 nm (420-1800 nm)) and NIR (λ> 780 nm (780-1800 nm) irradiation, respectively, compared with 0.3 and 0.10 μmol h-1 for BP NS/Pt (3 wt%)/P25. Moreover, a comparative study was made to examine the effect of thickness of BP NS on the photocatalytic H2 evolution. Femtosecond time-resolved diffused reflectance spectroscopy (fs-TRDRS) was integrated together with photoelectrochemical measurement to shed the light on the importance of charge transfer and separation, confirming that decreasing the thickness of BP NS enhances electron injection from BP NS to TMC to increase the photocatalytic activity.
关键词: hydrogen evolution,Black phosphorus,visible-light photocatalyst,charge carriers dynamics,TiO2 mesocrystals,femtosecond time-resolved diffuse reflectance
更新于2025-11-19 16:51:07
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Photo-physical properties of substituted 2,3-distyryl indoles: Spectroscopic, computational and biological insights
摘要: The structural dependence of the photo-physical properties of substituted 2,3-distyryl (23DSI) indoles were studied using several spectroscopic techniques including steady-state UV-VIS spectroscopy, steady-state fluorescence spectroscopy, steady-state excitation spectroscopy, time correlated single photon counting (TCSPC) spectroscopy, and time-resolved fluorescence upconversion spectroscopy (TRFLS). Each of 23DSI derivatives investigated showed distinct fluorescence emission and UV-VIS spectra, indicating strong structural dependence of the emission and the excitation. The UV-VIS spectra of the 23DSI derivatives showed three main identical absorption bands with minor deviations in the absorbance caused by substituent groups on the distyryl rings. The time-resolved fluorescence up-conversion studies indicated that the fluorescence undergoes a mono-exponential decay whereas the calculated fluorescence lifetime showed relatively short fluorescence lifetimes of approximately 1 ns. All of the 23DSI derivatives showed two-photon absorption upon direct excitation of 1.6 W laser pulses at 800 nm. These studies suggest that the substituents, attached to distyryl core, are capable of boosting or hindering fluorescence intensities by distorting the π-conjugation of the 23DSI molecule. Our studies showed that 23DSI (p-F) has the highest fluorescence emission quantum yield. Theoretical calculations for the ground state of 23DSI derivatives confirmed differences in electron densities in 23DSI derivatives in the presence of different substituent attachments. The excellent fluorescence emission, high fluorescence quantum yield and two-photon absorption properties of these 23DSI molecules make them attractive candidates for potential applications in the fields of biological imaging, biomedicine, fluorescent probes, and photodynamic inactivation (PDI). B. subtilis samples, treated with micro molar solutions of 23DSI (p-OCH3) and 23DSI (p-CH3), showed very effective photodynamic inactivation (PDI) upon irradiation with white light.
关键词: Two photon absorption,Time-resolved fluorescence upconversion laser spectroscopy,Photodynamic inactivation,Gaussian calculations,Photo-physical properties,Light-activation
更新于2025-11-14 15:26:12
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Dynamics of infrared excitations in boron doped diamond
摘要: We report on the investigation of relaxation dynamics of optical excitations in IIb high pressure high temperature (HPHT) diamond doped by natural boron and isotopically enriched 11boron. The measurements were performed with a pump-probe technique using short pump pulses from a wavelength-tunable infrared free electron laser. Lifetimes of excited boron states ranging from a few picoseconds to a few hundred picoseconds, have been derived from the obtained data. The relaxation rates depend on the pumped states, the pump intensity and the diamond lattice temperature. We discuss possible contributions to the optical and nonradiative intracenter relaxation rates observed in these experiments. Theoretical simulations support ultrafast relaxation by multiple phonon emission, for the electronic states with the energy gap exceeding the energy of optical phonon.
关键词: time-resolved spectroscopy,boron-doped diamond,diamond
更新于2025-11-14 15:14:40
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Extrinsic heavy metal atom effect on the solid state room temperature phosphorescence of cyclic triimidazole
摘要: Four coordination compounds [Zn3(CH3COO)6(H2O)2](TT)2, [Cd(H2O)6](ClO4)2(TT)2, [Cd(H2O)6](BF4)2(TT)2, [Zn(H2O)6](BF4)2(TT)2 (1-4) accommodating triimidazo[1,2-a:1',2'-c:1'',2''-e][1,3,5]triazine (TT) as a guest in their crystal lattice are isolated and fully photophysically and structurally characterized. Their emission properties are compared with those of afterglow TT and interpreted taking into account the heavy atom effect and crystal packing similarities and differences. In the case of 1, due to the closeness of the TT H-aggregates arrangement with that of the phosphor's pure phase, the observed intensification of the phosphorescent emission at the expense of the prompt one is attributed to the extrinsic heavy atom effect of Zn. In 2 and 3, the heavier Cd atom is responsible for a decrease in the lifetimes of the afterglow emission, despite the presence of tightly overlapped H-dimers in the crystal structure. Finally for 4, isostructural with 3, the Zn atom reveals in RTUP lifetime comparable with that of 1.
关键词: extrinsic heavy atom effect,H aggregates,time resolved photoluminescence,room temperature ultralong phosphorescence,single crystal XRD
更新于2025-09-23 15:23:52
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37ps-Precision Time-Resolving Active Quenching Circuit for High-Performance Single Photon Avalanche Diodes
摘要: Time-resolved imaging by means of single-photon avalanche diodes (SPADs) has achieved widespread interest in recent years, especially since technological progress has opened the way to the development of multichannel time-correlated single-photon counting (TCSPC) acquisition systems. Unfortunately, currently available TCSPC imagers feature relatively low performance with respect to state-of-the-art single-channel systems. A real breakthrough in this field would be the exploitation of large arrays of high-performance SPAD detectors developed by means of dedicated fabrication processes, usually referred to as custom technology. Custom-technology SPADs require external electronics potentially leading to interconnection issues for densely integrated arrays. In this paper, we present a new fully integrated front-end circuit able to provide both quenching/reset and timing functionalities while requiring a single connection toward the SPAD. This is the first fully integrated circuit reported in literature that can provide both the timing information about the photon time of arrival with a jitter as low as 37 ps and apply high-voltage pulses up to 50 V in order to meet the requirements of several detectors, including the new red-enhanced SPAD. Combining these two capabilities in a single circuit strongly reduces the complexity of the connection between an array of custom-technology SPADs and the relative external front end, thus paving the way for the exploitation of high-performance SPADs in TCSPC imaging systems.
关键词: Active Quenching Circuit,AQC,timing,time-resolved,SPAD,Single Photon Avalanche Diode,SPAD front end
更新于2025-09-23 15:23:52
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Smartphone-based apparatus for measuring upconversion luminescence lifetimes
摘要: Luminescence lifetime detection plays an important role in time-resolved detection and research. However, the traditional instruments always require expensive detectors such as time-correlated single photon counter or streak camera. Herein, a low-cost and miniaturized apparatus for measuring upconversion luminescence lifetimes was developed by using a smartphone equipped with a 980 nm CW laser and a motor. When the motor was driving the sample circling at a high linear velocity, the excited sample would emit a luminescence arc, which could be photographed by the phone camera. The rotating rate could be measured by a tuner APP and then used for transferring arc length to delay times. By analyzing the grayscale distribution of the luminescence arc, the luminescence decay curve was obtained, which was then used for exponential fit and calculating lifetimes. The whole system was not only remarkably cheaper but also more miniaturized than traditional instruments for measuring luminescence lifetimes, indicating the promising applications in point of care testing for time-resolved luminescence detection for bioanalysis and disease diagnosis.
关键词: Lanthanide upconversion,Luminescence lifetime,Time-resolved,Motor,Smartphone
更新于2025-09-23 15:23:52
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Effect of monatomic and molecular ion irradiation on time resolved photoluminescence decay in GaN
摘要: Optical effects induced in silicon-doped wurtzite (0001) GaN epilayers by keV monatomic and molecular ion irradiation were experimentally investigated. Results were analyzed together with data on structure defect formation. In all the cases under consideration, an increase in the collision cascade density (the cases of molecular and heavy atomic ion bombardment) enhances the stable damage accumulation rate and, accordingly, intensifies quenching of luminescence. The processes of PL suppression were theoretically considered as an increase of surface recombination rate of nonequilibrium photo-excited charge carriers due to production of stable damage at the irradiated subsurface layer. It is shown that carrier diffusion determines PL decay time shortening in the shallow implantation cases studied.
关键词: Radiation damage,Photoluminescence,Collision cascade density,Time-resolved PL,Charge carrier diffusion,Ion implantation,GaN
更新于2025-09-23 15:23:52
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Observation of Site-Resolved Vibrational Energy Transfer Using a Genetically Encoded Ultrafast Heater
摘要: Allosteric information transfer in proteins has been linked to distinct vibrational energy transfer (VET) pathways in a number of theoretical studies. Experimental evidence for such pathways, however, is sparse because site-selective injection of vibrational energy into a protein, i.e. localized heating, is required for their investigation. Here, we solve this problem by the site-specific incorporation of the non-canonical amino acid β-(1-azulenyl)-L-alanine (AzAla) via genetic code expansion. Being an exception to Kasha′s rule, AzAla undergoes ultrafast internal conversion and heating after S1 excitation while upon S2 excitation it serves as a fluorescent label. We endowed PDZ3, a protein interaction domain of postsynaptic density protein 95, with this ultrafast heater at two distinct positions. Using ultrafast IR spectroscopy, we could indeed observe VET from the incorporation sites in the protein to a bound peptide ligand on a picosecond timescale. This approach based on genetically encoded AzAla paves the way for detailed studies of VET and its role for function in a wide range of proteins.
关键词: protein modification,energy transfer,non-canonical amino acid,time-resolved spectroscopy,mutagenesis
更新于2025-09-23 15:23:52
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Role of solvent H-bonding and polarity on photophysical properties of a benzothiazole-based ratiometric amyloid fibril sensor
摘要: Detailed photophysical properties of 2-[2′-Me,4′-(dimethylamino)-phenyl]benzothiazole (2Me-DABT), a potential ratiometric amyloid fibril sensor, have been investigated in different solvents and solvent mixtures using steady state and time-resolved spectroscopic techniques. Our studies show that emission properties of 2Me-DABT are significantly modulated by polarity of solvent media. Extent of solvatochromism (3585 cm?1) shown by 2Me-DABT is quite large and can be used to monitor micropolarity of different complex media. Further, Stokes’ shift (> 10,000 cm?1) shown by 2Me-DABT in all studied solvents is also much larger than most common molecular probes. It has also been shown that hydrogen bonding with solvent molecules results in large modulation in photophysical properties of 2Me-DABT. The H-bonding with water molecules induces a large change (39°) in the dihedral angle between benzothiazole and aniline moieties resulting large changes in photophysical properties of 2Me-DABT in polar protic solvents. Photophysical properties reported herein are used to explain observed amyloid sensing behaviour of 2Me-DABT. Detailed quantum chemical calculations are performed to support the experimental results.
关键词: Amyloid sensor,H-bonding,Photophysics,Time-resolved spectroscopy,Fluorescence
更新于2025-09-23 15:23:52