Single Photon Counting Research Articles

Usefulness of a new anthropomorphic phantom simulating the chest and abdomen regions in PET tests.

To investigate the clinical utility of a new anthropomorphic phantom that reproduces the chest and abdomen better than the conventional National Electrical Manufacturers Association (NEMA) body phantom, count rates and image quality of PET images obtained from patients were evaluated. Anthropomorphic phantoms were used to include radioactivity in the lung, liver, kidney, and background regions. Two NEMA body phantoms were used for chest and abdominal assessments. The cross calibration factor (CCF) cylinder phantom was also used to reproduce the distribution of radioactivity outside the field of view, simulating the patient brain. Four types of phantoms were used in the PET imaging experiment, and for each phantom, the prompt coincidence count rates, random coincidence count rates, true + scatter coincidence count rates, and single photon count rates were measured. Then, these count rates were compared with count rates from actual clinical data. PET image quality assessment was done using the parameters, noise equivalent count patient (NECpatient), noise equivalent count density (NECdensity), and liver signal-to-noise ratio (SNR). Random coincidence count rates showed that the data obtained from each phantom were in good agreement with the clinical data. True + scatter coincidence count rates had better agreement with clinical data when measured for anthropomorphic phantoms than for the NEMA body phantoms. Furthermore, when the CCF Cylinder phantom simulating the brain was placed outside the imaging field of view, the results were closer to the clinical data. PET image quality was 1.4% higher for NECpatient obtained from anthropomorphic phantoms compared to the mean obtained from clinical data. NECdensity was 15.0% lower than the mean value obtained from clinical data. Liver SNR was 14.8% higher in PET images reconstructed using the 3D-ordered subsets expectation maximization (OSEM) method. It was 10.0% lower in PET images reconstructed with the image reconstruction method Q.Clear (GE Healthcare) using the Bayesian penalized likelihood (BPL) method. The new anthropomorphic phantom was more consistent with the count rates obtained from clinical data than the conventional NEMA body phantoms were and it was able to better simulate the distribution of radioactivity concentrations in the patients by reproducing the distribution of radioactivity concentrations outside the field of view.

The Molecular Mechanism of Fluorescence Lifetime of Fluorescent Probes in Cell Membranes.

Fluorescence probes play crucial roles in unraveling the structure and dynamics of cell membranes including membrane fluidity, polarity, and lipid molecule ordering. The fluorescence lifetime of probes describes the average duration of time that a fluorescent molecule remains in an excited state before returning to the ground state, which is sensitive to environmental changes. However, the molecular mechanism and inherent properties to determine the fluorescence lifetimes remain unexplored and inadequately studied. Furthermore, the effects of the probe on the membrane are also unclear. In this study, we investigated the interactions between probes and lipids, as well as the structural properties of probes within the outer and inner membrane of Mycobacterium smegmatis (Msm) by combining molecular dynamics (MD) simulations, enhanced sampling methods, fluorescence lifetime imaging microscopy (FLIM), and time-correlated single photon counting (TCSPC). The results show that even though the probes have very little effect on the membrane lipids, different membrane environments significantly affect the fluorescence lifetime of the probes. The analysis based on the all-atom simulations shows a strong correlation between the probe's immersion depth within the membrane and its fluorescence lifetime. Specifically, probes buried in the membrane environment shielded from rapid water molecule collisions exhibit longer fluorescence lifetimes. The molecular basis of the fluorescence lifetime of probes in cell membranes revealed in this work would enhance the comprehension of fluorescence probes and facilitate the rational design of novel efficient probes.

Exploring photophysical behavior and fullerene-induced quenching in difluoroboron flavanone [formula omitted]-diketonates for application in organic electronic devices: Experimental and theoretical analysis

The photophysical properties of two difluoroboron flavanone β-diketonates (DK1 and DK2) and their interaction with fullerene (C60) in toluene solution and spin-coated films were investigated using time-correlated single-photon counting, absorption spectroscopy, and steady-state fluorescence spectroscopy. In molecular crystal, the complexes exhibited red-shifted absorption and emission relative to their spectra in solution. Additionally, both complexes displayed bi-exponential decay behavior in time-resolved fluorescence measurements, indicating their capability to form both H and J types of aggregates in the solid state. The introduction of C60 resulted in significant fluorescence quenching and reduced excited-state lifetimes for both complexes. This quenching, observed in both solution and spin-coated films, was primarily driven by photo-induced electron transfer (PET) processes, underscoring the potential of these complexes as donors in fullerene-based heterojunction organic solar cells. To elucidate the process of aggregate formation and the impacts of different dimerization types within the crystalline structure of the complexes, first-principles calculations using Density Functional Theory (DFT) and time-dependent density functional theory (TD-DFT) were performed. We also employed DFT to explore various DK configurations on the fullerene surface, evaluating intermolecular distances and formation energies. These calculations highlighted the energetically favourable gap between the low-lying LUMO levels of the complexes and C60, confirming their suitability for such applications.

Blue Light Emitting Electron-Rich Triphenylamine Decorated Stilbene Derivatives for the Bio-Imaging Application

The stilbene functionalized star and bent shaped π-conjugate organic donor-donor (D-π-D) molecules with decorated triphenylamine central core were synthesized via Michaelis-Arbuzov and Wittig-Horner Reaction with excellent regio-specific. The photo-physical investigations clearly indicate the synthesized molecules have excellent optical properties with UV-visible regions and hypochromic shifts. The intra-molecular charge transfer (ICT) absorption of bent and star-shaped molecules was found in the region of 369–390 and 382–390 nm. The star and bent-shaped molecules exhibited emission maximums between 444–490 and 437–476 nm. While increasing the solvent polarity, the emission maximum of TDMSPA and DMSNDMPA molecules were observed in red-shift. The excited state dipole moment changed when compared to ground state due to the triphenylamine core confirmed by the Lippert-Mataga plot. The density functional theory is good evidence; the TDMSPA molecule has a twisted configuration and supports optical and electrochemical studies. Time-correlated single photon counting (TCSPC) technique showed that the excited state fluorescence lifetime of the stilbene molecules was found to be 2.25 and 2.18 ns for star and bent shaped molecules, respectively. The synthesized molecules exhibit significantly low band-gap energies between HOMO and LUMO, and the values are observed in the range between 2.86 and 2.91 eV. The triphenylamine core-based π-conjugate organic chromophore is widely used as a bio-imaging agent in Rhizoctonia oryzae (Plant fungal pathogen) as the model and the TDMSPA was an excellent bio-imaging agent and biocompatibility with fungal and bacterial cells.

Theoretical analysis of argon 2p states' density ratios for nanosecond plasma optical emission spectroscopy

A theoretical analysis of excited argon state densities responsible for optical emission spectra of atmospheric pressure argon plasma is presented for its use in plasma diagnostics. Nanosecond pulsed barrier discharges are simulated using spatially one- and two-dimensional fluid-Poisson models using the reaction kinetics model presented by Stankov et al. [1], which considers all ten argon 2p states (Paschen notation) separately. The very first (single) discharge and repetitive discharges with frequencies from 5 kHz to 100 kHz are considered. A semi-automated procedure is utilized to find appropriate 2p states for electric field determination using an intensity ratio method, which is based on a time-dependent collisional-radiative model. The fluid simulations in combination with the semi-automated procedure are used to quantify the sensitivity of selected 2p-state ratios to given preionization of the gas. A highly sensitive time-correlated single photon counting experiment shows clearly that the selected ratio is sensitive to the electric field variation in the streamer head, yet additional calibration is needed for absolute values determination. Different approaches for effective lifetime determination are tested and applied also to measured data. The influence of radial and axial 2p state density integration on the intensity ratio method is discussed. The above mentioned models and procedures result in a flexible theory-based methodology applicable for development of new diagnostic techniques.

Fine-structure changing collisions in 87Rb upon D2 excitation in the hyperfine Paschen-Back regime

Abstract We investigate fine structure changing collisions in 87Rb vapour upon D2 excitation in a thermal vapour at 75 ∘C; the atoms are placed in a 0.6 T axial magnetic field in order to gain access to the hyperfine Pashen-Back regime. Following optical excitation on the D2 line, the exothermic transfer 5P 3 / 2 → 5P 1 / 2 occurs as a consequence of buffer-gas collisions; the 87Rb subsequently emits a photon on the D1 transition. We employ single-photon counting apparatus to monitor the D1 fluorescence, with an etalon filter to provide high spectral resolution. By studying the D1 fluorescence when the D2 excitation laser is scanned, we see that during the collisional transfer process the m J ′ quantum number of the atom changes, but the nuclear spin projection quantum number, m I ′ , is conserved. A simple kinematic model incorporating a coefficient of restitution in the collision accounted for the change in velocity distribution of atoms undergoing collisions, and the resulting fluorescence lineshape. The experiment is conducted with a nominally ‘buffer-gas free’ vapour cell; our results show that fine structure changing collisions are important with such media, and point out possible implications for quantum-optics experiments in thermal vapours producing entangled photon pairs with the double ladder configuration, and solar physics magneto-optical filters.

Room temperature single-photon terahertz detection with thermal Rydberg atoms

Single-photon terahertz (THz) detection is one of the most demanding technologies for a variety of fields and could lead to many breakthroughs. Although significant progress has been made in the past two decades, operating it at room temperature still remains a great challenge. Here, we demonstrate, for the first time, a room temperature THz detector at single-photon levels based on nonlinear wave mixing in thermal Rydberg atomic vapor. The low-energy THz photons are coherently upconverted to high-energy optical photons via a nondegenerate Rydberg state involved in a six-wave mixing process, and therefore, single-photon THz detection is achieved by a conventional optical single-photon counting module. The noise equivalent power of such a detector reaches 9.5 × 10−19 W/Hz1/2, which is more than four orders of magnitude lower than the state-of-the-art room temperature THz detectors. The optimum quantum efficiency of the whole-wave mixing process is about 4.3%, with 40.6 dB dynamic range, and the maximum conversion bandwidth is 172 MHz, which is all-optically controllable. The developed fast and continuous-wave single-photon THz detector at room temperature operation has a great potential for portability and chip-scale integration, and could be revolutionary for a wide range of applications in remote sensing, wireless communication, biomedical diagnostics, and quantum optics.

MKIDGen3: Energy-resolving, single-photon-counting microwave kinetic inductance detector readout on a radio frequency system-on-chip.

Microwave Kinetic Inductance Detectors (MKIDs) are superconducting detectors capable of single-photon counting with energy resolution across the ultraviolet, optical, and infrared (UVOIR) spectrum with microsecond timing precision. MKIDs are also multiplexable, providing a feasible way to create large-format, cryogenic arrays for sensitive imaging applications in biology, astronomy, and quantum information. Building large, cryogenic MKID arrays requires processing highly multiplexed, wideband readout signals in real time; this task has previously required large, heavy, and power-intensive custom electronics. In this work, we present the third-generation UVOIR MKID readout system (Gen3), which is capable of reading out twice as many detectors with an order of magnitude lower power, weight, volume, and cost-per-pixel as compared to the previous system. Gen3 leverages the Xilinx RFSoC4x2 platform to read out 2048, 1MHz MKID channels per board. The system takes a modern approach to FPGA design using Vitis High-Level Synthesis to specify signal processing blocks in C/C++, Vivado ML intelligent design runs to inform implementation strategy and close timing, and Python productivity for Zynq to simplify interacting with and programming the FPGA using Python. This design suite and tool flow allows general users to contribute to and maintain the design and positions Gen3 to rapidly migrate to future platforms as they become available. In this work, we describe the system requirements, design, and implementation. We also provide performance characterization details and show that the system achieves detector-limited resolving power in the case of few readout tones and minimal degradation with all 2048 tones. Planned upgrades and future work are also discussed. The Gen3 MKID readout system is fully open-source and is expected to facilitate future array scaling to megapixel-sized formats and increase the feasibility of deploying UVOIR MKIDs in space.

Uncovering Integrated Dual-State ESIPT-Conductivity, Redox-Capacity, and Opto-Electronic Responses Toward Hg(II)/ Cr(III) of Aliphatic Fluorescent Polymers.

Excited-state intramolecular proton transfer (ESIPT)-associated dual-state emissive aliphatic dual-light emitting conducting polymers (DLECPs) having oxidation-reduction capacities are prepared polymerizing 2-acrylamido-2-methylpropane-1-sulfonic acid, methacrylic acid, and 2-methyl-3-(N-(2-methyl-1-sulfopropan-2-yl)acrylamido)propanoic acid monomers. Of as-synthesized DLECPs, nuclear magnetic resonance (NMR) and Fourier transform infrared (FTIR) spectroscopies, fluorescent enhancements (I/I0), and computational investigation indicate intriguing photophysical features in DLECP3 (optimum composition). In DLECP3, ─CONH─, ─CON<, and ─COOH subluminophores are recognized by density-functional theory (DFT)/time-dependent-DFT calculations and experimental investigations. ESIPT-associated dual-state emission/conductivity, aggregation-enhanced emissions, selective opto-electronic responses toward Hg(II)/Cr(III) at 437/574nm, and redox properties of DLECP3 are supported by solid-state/solution spectroscopies, time-correlated single photon counting (TCSPC) measurements, dual-state excitation dependent emissions, microscopic images, electrochemical measurements, and DFT calculations. Here, preferential interaction of Hg(II)/Cr(III) with DLECP3 (amide)/DLECP3 (imidol) and reduction/oxidation of Hg(II)/Cr(III) to Hg(I)/Cr(VI) are substantiated by UV-vis, FTIR, and X-ray photoelectron spectroscopies; TCSPC measurements; NMR-titration; electrochemical studies; alongside computational calculations. The proton-electrical conductivities of DLECP3, Hg(II/I)-DLECP3, and Cr(III/VI)-DLECP3 in solids/solutions are 15.27 × 10-5/6.16 × 10-5, 19.60 × 10-5/25.52 × 10-5, and 26.69 × 10-5/27.60 × 10-5 S cm-1, respectively.

Nonclassical Correlations between Photons and Phonons of Center-of-Mass Motion of a Mechanical Oscillator.

We demonstrate nonclassical correlations between phonons and photons created using opto-mechanical spontaneous parametric down-conversion in a system based on a soft-clamped ultracoherent membrane oscillator inside of a Fabry-Pérot optical resonator. Non-Gaussian quantum features are demonstrated for the center-of-mass motion of a submillimeter nanogram-scale mechanical oscillator. We show that phonons stored in the mechanical oscillator, when subsequently read out, display strong signs of quantum coherence, which we demonstrate by single-photon counting enabled by our state-of-the-art optical filtering system. We observe a violation of the classical two-time Cauchy-Schwarz inequality between a heralding write photon and a stored phonon with a confidence of >92%.

Temperature-dependent Rotational Dynamics of a 3-(benzo[d]Thiazol-2-yl)-7-diethylamino)-2H-chromen-2-one non-polar Probe in n-pentanol, n-nonanol, and n-decanol Solvents.

Rotational dynamics of non-polar probe 3-(benzo[d]thiazol-2-yl)-7-(diethylamino)-2H-chromen-2-one (3BT7D2H-one) and in n-pentanol, n-nonanol, and n-decanol alcohol solvents is studied using steady-state fluorescence depolarization and time-correlated single photon counting technique with varying temperature. Experimental observations indicate that 3BT7D2H-one rotates slowly in n-decanol solvent compared to other chosen solvents. Rotational motion is analyzed using the Stoke 's-Einstein-Debye (SED) model. 3BT7D2H-one solute rotational dynamics is satisfactorily described by the SED slip boundary limit in all three solvents.

Quantitative analysis of transferrin uptake into living cells at single-molecule level

Endocytosis, apart from providing a mechanism for cells to take up nutrients, is the principal route of entry for many nanomedicines. The evaluation of therapeutic delivery efficiency without providing quantitative parameters, like the number of molecules entering the cell and the time of their entry, is very limited. Despite advances in single-molecule methods for in-cell quantitative measurements, they have not become widely used in the study of endocytosis. Their application, however, is challenging owing to the appropriate experimental design and applicable analysis methods. Here, by using an integrated approach based on the multidimensional time-correlated single-photon counting (TCSPC) technique, we demonstrate the quantitative method to measure the time- and concentration-dependent uptake of TRITC-transferrin into living cells with single-molecule sensitivity. The methodology opens possibilities for quantitatively assessing the delivery efficiency of fluorescent molecules into living cells.

Quantitative Analysis of Acquisition Speed of High-Precision FLIM Technologies via Simulation and Modeling

In practical applications such as cancer diagnosis and industrial detection, there is a critical demand for fast fluorescence lifetime imaging (Fast-FLIM). The Fast-FLIM systems suitable for complex environments are typically achieved by enhancing the hardware performance of time-correlated single-photon counting (TCSPC), with an acquisition speed of about a few frames per second (fps). However, due to the limitation of single-photon acquisition, the imaging speed is still far from the demand of practical application. The synchroscan streak camera (SC) maps signals from the temporal dimension to the spatial dimension, effectively overcoming the long acquisition time caused by single-photon acquisition. This paper constructs a method to calculate the acquisition time for the TCSPC-FLIM and SC-FLIM systems, and it quantitatively compares the speed. The research demonstrates that the main factors limiting the acquisition speed of the FLIM systems are the photon emission rate, the photon counting rate, the required SNR, the dwell time, and the number of parallel channels. In high-quality and large-scale lifetime imaging, the acquisition speed of the SC-FLIM is at least 104 times faster than that of the TCSPC-FLIM. Therefore, the synchroscan streak camera has more significant potential to promote Fast-FLIM.

Surface roughness metrology with a raster scanning single photon LiDAR

We explore a novel, to the best of our knowledge, approach to surface roughness metrology utilizing a single pixel, raster scanning single photon counting LiDAR system. It uses a collimated laser beam in picosecond pulses to probe a surface, capturing the changes of back-scattered photons from different points on the surface into a single mode fiber, and counting them using a single photon detector. These back-scattered photons carry speckle noise produced by the rough surface, and the variation in photon counts over different illumination points across the surface becomes a good measure of its roughness. By analyzing the variation frequency as the LiDAR scans over the surface using machine learning techniques, we demonstrate general measurements of surface roughness from 1.21 (1.27±4.51) to 102.01 (87.97±10.55) microns.

Exploring Through-Space Charge Transfer-Mediated Optoelectrochemical Properties of Dual-State Luminescent Aliphatic Polymers and Optoelectronic Responses toward Metal Ions.

Herein, natural-synthetic hybrid dual-state luminescent conducting polymers (DLCPs/DLCP1-DLCP8) possessing significant optoelectrochemical properties are strategically developed by the polymerization of prop-2-enamide, cis-butenedioic acid, 2-acrylamido-2-methylpropane-1-sulfonic acid, and in situ-generated 2-(3-acrylamidopropanamido)-2-methylpropane-1-sulfonic acid alongside the grafting of gum tragacanth. The spectroscopic data of aliphatic DLCPs affirm DLCP7 as the most stable supramolecular assembly endowing optoelectronic properties. Computational calculations identified -C(═O)NH-, -C(═O)OH, -OH, and -SO3H as subluminophores. The absorption spectra, excitation wavelength-/solvent-polarity-/concentration-dependent luminescence, solid state luminescence, aggregation-induced enhanced luminescence, and time-correlated single photon count (TCSPC) studies confirm the occurrence of aggregation-mediated intramolecular through-space charge transfer (ITSCT) in the excited state of DLCP7. Mulliken charge, natural bond orbital, dipole moments, and electronic potential surface analyses confirm the charge donor-acceptor system in DLCP7. Furthermore, the selective optoelectronic response of DLCP7 toward Ca2+/Cu(II) at 438/574 nm is explored using ultraviolet-visible spectra, TCSPC analyses, a dynamic light scattering study, and computational investigations. The chelation-enhanced luminescence and ITSCT inhibition are responsible for turn-on and turn-off detections of Ca2+ and Cu(II), respectively. Cu(II) → Cu(I) reduction in a DLCP7 solution is inferred from electrochemical and spectroscopic analyses. The conductivities of 9.65 × 10-5 S cm-1 (solid state) and 44.35 × 10-5 S cm-1 (solution) in DLCP7 are validated by current-voltage and electrochemical impedance measurements. Again, strong electronic conductivities of 43.89 × 10-5 S cm-1 (solid state)/53.34 × 10-5 S cm-1 (solution) and 45.42 × 10-5 S cm-1 (solid state)/64.81 × 10-5 S cm-1 (solution) are observed in Ca2+-DLCP7 and Cu(II)-DLCP7, respectively.

Storehouse of Peculiar Supramolecular Architecture: Probing the Charge-Transfer Phenomenon and Effect of Altering the pH in a Meta-Oriented Single Donor-Double Acceptor Fluorophore.

The investigation of excited-state intramolecular charge transfer (ESICT) has been a fascinating area of research. Although the ESICT events have been studied mostly for para-disubstituted donor-acceptor type molecules, the meta-oriented donor-acceptor type molecules have also shown tremendous potential as ESICT active molecules. In the current work, a small fluorescent probe diethyl 5-amino isophthalate (DE-5A-IPA) was investigated as a potential model to investigate ESICT events in the solution as well as solid phase. DE-5A-IPA was synthesized easily starting from commercially available 5-amino isophthalic acid in excellent yield. In the solid state, differing extents of CH···π-type binding led to formation of fully planar and puckered "cyclobutane" mimic supramolecular architecture, as evidenced from the crystal structure analysis. In addition, the single crystal structure of DE-5A-IPA shows that the donor (-NH2) and acceptor (-CO2Et) are situated in the same plane, thereby assuring the prerequisites of a through-space charge transfer. DE-5A-IPA undergoes noticeable Stokes shift (in cm-1) when the polarity of the medium was changed (4820 cm-1 in hexane; 9375 cm-1 in water). The excited-state dipole moment (μe) was calculated to be 4.0 units higher than the ground-state dipole moment (μg). DE-5A-IPA had an appreciable quantum yield (0.10 ± 0.03 to 0.27 ± 0.04). The ESICT phenomenon was also investigated by ground- and excited-state structural calculations using Gaussian 16 software. The excited-state lifetime measured by the time correlated single photon counting technique was found to vary with the polarity of the solvent, thereby providing further support to the ESICT phenomenon being operative in DE-5A-IPA. Taking advantage of the -NH2 group (which is susceptible to protonation) in DE-5A-IPA, steady state and time-resolved photodynamics were investigated in solutions of varying pH values. Interestingly, the emission quantum yields as well as the emission lifetime increase with an increase in pH value, thereby establishing DE-5A-IPA as a pH-sensitive probe. The current findings shall boost the understanding of meta-oriented ESICT enabled compounds in terms of their excited-state photophysics.

The use of low-frequency current fluctuations in measuring the mobility of holes in the MEH-PPV polymer

A diagnostic method based on the evaluation of low-frequency current fluctuation spectra is presented. When measuring the current through a p-type MEH-PPV sample, the occurrence of fluctuation is observable which can be measured with an AC amplifier. A model has been proposed that proves that the fluctuations originate from the interaction between the valence band and the band gap traps. The mean value of the amplitudes of these fluctuations increases linearly with decreasing frequency with a slope from which the product of mobility and lifetime of current carriers µpτp= (9 ± 3) × 10−15 cm2V−1 was obtained. The hole lifetime of (0.27 ± 0.01) ns was evaluated from the luminescence relaxation using the time-correlated single photon counting (TCSPC) technique. The mobility value (3 ± 1) × 10−5 cm2 V−1s−1 calculated using the above methods was compared with the mobility 1.8 × 10−5 cm2 V−1s−1 determined by the CELIV method (Charge extraction by linearly increasing voltage) and good agreement was obtained.

MoS2-CZTS: a 2D-3D nanocomposite to enhance the photocatalytic performance in degradation of methylene blue dye

Cu2ZnSnS4 nanocrystals (CZTS NCs), MoS2 nanosheet (NS) and MoS2-CZTS nanocomposite (NC) have been synthesized using solvothermal route. Structural characterization of samples have been done by XRD, Raman spectroscopy, HR-TEM. Samples have optically characterized by UV-Vis absorption, photoluminescence (PL) and time co-related single photon counting (TCSPC) study. The XRD, HR-TEM and Raman spectroscopy established tetragonal kesterite phase for both CZTS NCs and MoS2-CZTS NC. Enhancement of efficiency of CZTS NCs to degrade methylene blue (MB) dye, illuminated by visible light, have been observed by loading 1 wt.% MoS2 NS and found to be ~100% in only 15 minutes. This is due to efficent transfer of charge carriers at p-CZTS and n-MoS2 heterojunction interface, confirmed by quenching of PL intensity and decrease in average lifetime of carriers.

Synthesis and optimization of chitosan-incorporated semisynthetic polymer/α-Fe2O3 nanoparticle hybrid polymer to explore optimal efficacy of fluorescence resonance energy transfer/charge transfer for Co(II) and Ni(II) sensing

Initially, four synthetic fluorescent polymers (SFPs) are synthesized from α-methacrylic acid and methanolacrylamide monomers carrying –C(=O)OH and –C(=O)NH subfluorophores, respectively. Among SFPs, ∼1:1 incorporation of subfluorophores in the optimum SFP3 is explored by spectroscopic analyses. Subsequently, chitosan is incorporated in SFP3 to produce five semi-synthetic fluorescent polymers (SSFPs). The maximum incorporation of chitosan in SSFP4 is supported by different spectroscopies. In SSFP4, strong electrostatic interactions among polar functionalities of chitosan and synthetic polymer favor resonance-associated charge transfer (RCT) from SSFP4-(amide) to SSFP4-(canonical). Finally, three hybrid fluorescent polymers (HFPs) are fabricated encapsulating iron-oxide nanoparticle within SSFP4. The maximum proportion of hematite (α-Fe2O3) phase in HFPs is explored by spectroscopic, magnetometric, microscopic, and light scattering studies. HFP2 shows local/RCT/fluorescence resonance energy transfer (FRET) emission at 393/460/570 nm. In HFP2, FRET, RCT, and ratiometric pH-sensing within 3.0–6.5 phenomena are explored by solvent polarity effects, time-correlated single photon counting, quantum yield measurements, alongside I431/I460 vs pH plots. RCT and FRET emissions of HFP2 are utilized for selective sensing of Co(II)/Ni(II) with limits of detection of 4.990 ppb (460 nm)/4.353 ppb (570 nm) and 45.041 ppb (428 nm)/29.617 ppb (527 nm) in organic and aqueous solutions, respectively.

Fluorescence-Detected Two-Dimensional Electronic Spectroscopy of a Single Molecule.

Single-molecule fluorescence spectroscopy is a powerful method that avoids ensemble averaging, but its temporal resolution is limited by the fluorescence lifetime to nanoseconds at most. At the ensemble level, two-dimensional spectroscopy provides insight into ultrafast femtosecond processes, such as energy transfer and line broadening, even beyond the Fourier limit, by correlating pump and probe spectra. Here, we combine these two techniques and demonstrate coherent 2D spectroscopy of individual dibenzoterrylene (DBT) molecules at room temperature. We excite the molecule in a confocal microscope with a phase-modulated train of femtosecond pulses and detect the emitted fluorescence with single-photon counting detectors. Using a phase-sensitive detection scheme, we were able to measure the nonlinear 2D spectra of most of the DBT molecules that we studied. Our method is applicable to a wide range of single emitters and opens new avenues for understanding energy transfer in single quantum objects on ultrafast time scales.