ABSTRACT
Developing deep learning models for accurate segmentation of biomedical CT images is challenging due to their complex structures, anatomy variations, noise, and unavailability of sufficient labeled data to train the models. There are many models in the literature, but the researchers are yet to be satisfied with their performance in analyzing biomedical Computed Tomography (CT) images. In this article, we pioneer a deep quasi-recurrent self-attention structure that works with a dual encoder-decoder. The proposed novel deep quasi-recurrent self-attention architecture evokes parameter reuse capability that offers consistency in learning and quick convergence of the model. Furthermore, the quasi-recurrent structure leverages the features acquired from the previous time points and elevates the segmentation quality. The model also efficiently addresses long-range dependencies through a selective focus on contextual information and hierarchical representation. Moreover, the dynamic and adaptive operation, incremental and efficient information processing of the deep quasi-recurrent self-attention structure leads to improved generalization across different scales and levels of abstraction. Along with the model, we innovate a new training strategy that fits with the proposed deep quasi-recurrent self-attention architecture. The model performance is evaluated on various publicly available CT scan datasets and compared with state-of-the-art models. The result shows that the proposed model outperforms them in segmentation quality and training speed. The model can assist physicians in improving the accuracy of medical diagnoses.
ABSTRACT
An introduction to the joint Nanoscale and Chemical Communications (ChemComm) themed collection focused on fundamental processes in optical nanomaterials that features a series of articles describing the properties of this versatile class of materials while highlighting some of their potential applications.
ABSTRACT
When misfolded, α-Synuclein (α-Syn), a natively disordered protein, aggregates to form amyloid fibrils responsible for the neurodegeneration observed in Parkinson's disease. Structural studies revealed distinct molecular packing of α-Syn in different fibril polymorphs and variations of interprotofilament connections in the fibrillar architecture. Fibril polymorphs have been hypothesized to exhibit diverse surface polarities depending on the folding state of the protein during aggregation; however, the spatial variation of surface polarity in amyloid fibrils remains unexplored. To map the local polarity (or hydrophobicity) along α-Syn fibrils, we visualized the spectral characteristics of two dyes with distinct polarities-hydrophilic Thioflavin T (ThT) and hydrophobic Nile red (NR)âwhen both are bound to α-Syn fibrils. Dual-channel fluorescence imaging reveals uneven partitioning of ThT and NR along individual fibrils, implying that relatively more polar/hydrophobic patches are spread over a few hundred nanometers. Remarkably, spectrally resolved sensitized emission imaging of α-Syn fibrils provides unambiguous evidence of energy transfer from ThT to NR, implying that dyes of dissimilar polarity are in close proximity. Furthermore, spatially resolved fluorescence spectroscopy of the solvatochromic probe NR allowed us to quantitatively map the range and variation of the polarity parameter ET30 along individual fibrils. Our results suggest the existence of interlaced polar and nonpolar nanoscale domains throughout the fibrils; however, the relative populations of these patches vary considerably over larger length scales likely due to heterogeneous packing of α-Syn during fibrilization and dissimilar exposed polarities of polymorphic segments. The employed method may provide a foundation for imaging modalities of other similar structurally unresolved systems with diverse hydrophobic-hydrophilic topology.
Subject(s)
Parkinson Disease , alpha-Synuclein , Humans , alpha-Synuclein/metabolism , Amyloid/metabolism , Parkinson Disease/metabolism , Optical Imaging , Coloring AgentsABSTRACT
Liquid-liquid phase separation (LLPS) has emerged as an important phenomenon associated with formation of membraneless organelles. Recently, LLPS has been shown to act as nucleation centers for disease-associated protein aggregation and amyloid fibril formation. Phase-separated α-synuclein droplets gradually rigidify during the course of protein aggregation, and it is very challenging to understand the biomolecular interactions that lead to liquid-like to solid-like transition using conventional ensemble measurements. Here, we describe a spectrally-resolved fluorescence microscopy based Förster resonance energy transfer (FRET) imaging to probe interactions of α-synuclein in individual droplets during LLPS-mediated aggregation. By acquiring entire emission spectral profiles of individual droplets upon sequential excitation of acceptors and donors therein, this technique allows for the quantification of sensitized emission proportional to the extent of FRET, which enables interrogation of the evolution of local interactions of donor-/acceptor-labeled α-synuclein molecules within each droplet. The present study on single droplets is not only an important development for studying LLPS but can also be used to investigate self-assembly or aggregation in biomolecular systems and soft materials.
Subject(s)
Fluorescence Resonance Energy Transfer , alpha-Synuclein , Fluorescence Resonance Energy Transfer/methods , Protein Aggregates , Microscopy, FluorescenceABSTRACT
Excitation energy migration beyond mesoscale is of contemporary interest for both solar photovoltaic and light-emissive devices, especially in context of organometal halide perovskites (OMHPs) which have been shown to have very long (charge carrier) diffusion lengths. While understanding the energy propagation pathways in OMHPs is crucial for further advancement of material design and improvement of opto-electronic features, the simultaneous existence of multiple processes like carrier diffusion, photon recycling, and photon transport makes it often complex to differentiate them. In this study, we unravel the diverse yet dominant excitation energy transfer mode(s) in crystalline MAPbBr3micron-sized 1D rods and plates by localized (confocal) laser excitation coupled with spectrally-resolved wide-field fluorescence imaging. While rarely used, this technique can efficiently probe excitation migration beyond the diffraction limit and can be realized by simple modification of existing epifluorescence microscopy setups. We find that in rods of length below â¼2 microns, carrier diffusion dominates amongst various energy transfer processes. However, the transient non-radiative defects severely inhibit the extent of carrier migration and also temporarily affect the radiative recombination dynamics of the photo-carriers. For MAPbBr3plates of several tens of micrometers, we find that the photoluminescence (PL) spectral characteristics remain unaltered at short distances (< â¼3µm) while at a larger distance, the spectral profile is gradually red-shifted. This implies that carrier diffusion dominates over small distances, while photon recycling,i.e., repeated re-absorption and re-emission of photons, propagates excitation energy transfer over extended length scales with assistance from wave-guided photon transport. Our findings can potentially be used for future studies on the characterization of energy transport mechanisms in semiconductor solids as well as for organic (molecular) self-assembled microstructures.
ABSTRACT
Amyloid fibrils are structurally heterogeneous protein aggregates that are implicated in a wide range of neurodegenerative and other proteopathic diseases. These fibrils exist in a variety of different tertiary and higher-level structures, and this exhibited polymorphism greatly complicates any structural study of amyloid fibrils. In this work, we demonstrate a method of using polarization-resolved microscopy to directly observe the structural heterogeneity of individual amyloid fibrils using amyloid-bound fluorophores. We formulate a mathematical quantity, helical anisotropy, which utilizes the polarized emission of amyloid-bound fluorophores to report on the local structure of individual fibrils. Using this method, we show how model amyloid fibrils generated from short peptides exhibit diverse structural properties both between different fibrils and within a single fibril, in a manner that is replicated for fibrils assembled from longer proteins. Our method represents an accessible and easily adaptable technique by which polymorphism in the structure of amyloid fibrils can be probed. Additionally, the methodology we describe here can be easily extended to the study of other fibrillar and otherwise ordered supramolecular structures.
Subject(s)
Amyloid , Peptides , Amyloid beta-Peptides , Microscopy, PolarizationABSTRACT
Aromatics such as phenols, benzene, and toluene are carcinogenic xenobiotics which are known to pollute water resources. By employing synthetic biology approaches combined with a structure-guided design, we created a tunable array of whole-cell biosensors (WCBs). The MopR genetic system that has the natural ability to sense and degrade phenol was adapted to detect phenol down to â¼1 ppb, making this sensor capable of directly detecting phenol in permissible limits in drinking water. Importantly, by using a single WCB design, we engineered mutations into the MopR gene that enabled generation of a battery of sensors for a wide array of pollutants. The engineered WCBs were able to sense inert compounds like benzene and xylene which lack active functional groups, without any loss in sensitivity. Overall, this universal programmable biosensor platform can be used to create WCBs that can be deployed on field for rapid testing and screening of suitable drinking water sources.
Subject(s)
Biosensing Techniques , Drinking Water , Environmental Pollutants , Benzene/analysis , Environmental Pollutants/analysis , XylenesABSTRACT
Tracking the movement of fluorescent single-molecule (SM) tracers has provided several new insights into the local structure and dynamics in complex environments such as soft materials and biological systems. However, SM tracking (SMT) remains unreliable at molecular length scales, as the localization error (LE) of SM trajectories (â¼30-50 nm) is considerably larger than the size of molecular tracers (â¼1-2 nm). Thus, instances of tracer (im)mobility in heterogeneous media, which provide indicators for underlying anomalous-transport mechanisms, remain obscured within the realms of SMT. Since the translation of passive tracers in an isotropic media is associated with fast dipolar rotation, we propose that authentic pauses within the LE can be revealed by probing the hindrance of SM reorientational dynamics. Here, we demonstrate how polarization-resolved SMT (PR-SMT) can provide emission anisotropy at each super-localized position, thereby revealing the tumbling propensity of SMs during random walks. For rhodamine 6G tracers undergoing heterogeneous transport in a hydrated polyvinylpyrrolidone (PVP) network, analysis of PR-SMT trajectories enabled us to discern instances of genuine immobility and localized motion within the LE. Our investigations on 100 SMs in (plasticized) PVP films reveal a wide distribution of dwell times and pause frequencies, demonstrating that most probes intermittently experience complete translational and rotational immobilization. This indicates that tracers serendipitously encounter compact, rigid polymer cavities during transport, implying the existence of nanoscale glass-like domains sparsely distributed in a predominantly deep-rubbery polymer network far above the glass transition.
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The study of controlling the morphology for designing advanced supramolecular architectures by tuning the molecular motif at the elemental level has been rarely carried out. Here, we report the synthesis of a nicotinic acid-conjugated selenopeptide, which induced the formation of an unbranched mesoscale elongated tubular morphology. We rationally designed two additional peptides to find out the decisive role played by the nitrogen atom (in nicotinic acid) and selenium (in the peptide backbone) toward the formation of the mesotube. We found that the peptide, devoid of nitrogen, forms a fibrillar structure, whereas the peptide without selenium self-assembled into a cylindrical filled rodlike morphology. Here, we report an entirely different class of peptide inspired from the selenopeptide chemistry that forms a tubular structure and unambiguously establish that both nicotinic acid and selenium are essential toward the formation of such mesotubes.
Subject(s)
Biocompatible Materials/chemistry , Niacin/chemistry , Peptides/chemistry , Selenium Compounds/chemistry , Biocompatible Materials/chemical synthesis , Materials Testing , Molecular Structure , Particle SizeABSTRACT
α-Synuclein (α-Syn) aggregation and amyloid formation is directly linked with Parkinson's disease pathogenesis. However, the early events involved in this process remain unclear. Here, using the in vitro reconstitution and cellular model, we show that liquid-liquid phase separation of α-Syn precedes its aggregation. In particular, in vitro generated α-Syn liquid-like droplets eventually undergo a liquid-to-solid transition and form an amyloid hydrogel that contains oligomers and fibrillar species. Factors known to aggravate α-Syn aggregation, such as low pH, phosphomimetic substitution and familial Parkinson's disease mutations, also promote α-Syn liquid-liquid phase separation and its subsequent maturation. We further demonstrate α-Syn liquid-droplet formation in cells. These cellular α-Syn droplets eventually transform into perinuclear aggresomes, the process regulated by microtubules. This work provides detailed insights into the phase-separation behaviour of natively unstructured α-Syn and its conversion to a disease-associated aggregated state, which is highly relevant in Parkinson's disease pathogenesis.
Subject(s)
Protein Aggregates/physiology , alpha-Synuclein/chemistry , HeLa Cells , Humans , Hydrogen-Ion Concentration , Microscopy, Confocal , Mutagenesis, Site-Directed , Parkinson Disease/metabolism , Parkinson Disease/pathology , Phase Transition , Polyethylene Glycols/chemistry , Recombinant Proteins/biosynthesis , Recombinant Proteins/chemistry , Recombinant Proteins/isolation & purification , alpha-Synuclein/genetics , alpha-Synuclein/metabolismABSTRACT
Supramolecular block copolymerzation with optically or electronically complementary monomers provides an attractive bottom-up approach for the non-covalent synthesis of nascent axial organic heterostructures, which promises to deliver useful applications in energy conversion, optoelectronics, and catalysis. However, the synthesis of supramolecular block copolymers (BCPs) constitutes a significant challenge due to the exchange dynamics of non-covalently bound monomers and hence requires fine microstructure control. Furthermore, temporal stability of the segmented microstructure is a prerequisite to explore the applications of functional supramolecular BCPs. Herein, we report the cooperative supramolecular block copolymerization of fluorescent monomers in solution under thermodynamic control for the synthesis of axial organic heterostructures with light-harvesting properties. The fluorescent nature of the core-substituted naphthalene diimide (cNDI) monomers enables a detailed spectroscopic probing during the supramolecular block copolymerization process to unravel a nucleation-growth mechanism, similar to that of chain copolymerization for covalent block copolymers. Structured illumination microscopy (SIM) imaging of BCP chains characterizes the segmented microstructure and also allows size distribution analysis to reveal the narrow polydispersity (polydispersity index (PDI) ≈ 1.1) for the individual block segments. Spectrally resolved fluorescence microscopy on single block copolymerized organic heterostructures shows energy migration and light-harvesting across the interfaces of linearly connected segments. Molecular dynamics and metadynamics simulations provide useful mechanistic insights into the free energy of interaction between the monomers as well as into monomer exchange mechanisms and dynamics, which have a crucial impact on determining the copolymer microstructure. Our comprehensive spectroscopic, microscopic, and computational analyses provide an unambiguous structural, dynamic, and functional characterization of the supramolecular BCPs. The strategy presented here is expected to pave the way for the synthesis of multi-component organic heterostructures for various functions.
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In layered hybrid perovskites, such as (BA)2 PbI4 (BA=C4 H9 NH3 ), electrons and holes are considered to be confined in atomically thin two dimensional (2D) Pb-I inorganic layers. These inorganic layers are electronically isolated from each other in the third dimension by the insulating organic layers. Herein we report our experimental findings that suggest the presence of electronic interaction between the inorganic layers in some parts of the single crystals. The extent of this interaction is reversibly tuned by intercalation of organic and inorganic molecules in the layered perovskite single crystals. Consequently, optical absorption and emission properties switch reversibly with intercalation. Furthermore, increasing the distance between inorganic layers by increasing the length of the organic spacer cations systematically decreases these electronic interactions. This finding that the parts of the layered hybrid perovskites are not strictly electronically 2D is critical for understanding the electronic, optical, and optoelectronic properties of these technologically important materials.
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Single molecule tracking experiments inside a hydrated polymer network have shown that the tracer motion is subdiffusive due to the viscoelastic environment inside the gel-like network. This property can be related to the negative autocorrelation of the instantaneous displacements at short times. Although the displacements of the individual tracers exhibit Gaussian statistics, the displacement distribution of all the trajectories combined from different spatial locations of the polymer network exhibits a non-Gaussian distribution. Here, we analyze many individual tracer trajectories to show that the central portion of the non-Gaussian distribution can be well approximated by an exponential distribution that spreads sublinearly with time. We explain all these features seen in the experiment by a generalized Langevin model for an overdamped particle with algebraically decaying correlations. We show that the degree of non-Gaussianity can change with the extent of heterogeneity, which is controlled in our model by the experimentally observed distributions of the motion parameters.
ABSTRACT
Photo-luminescence (P-L) intermittency (or blinking) in semiconductor nanocrystals (NCs), a phenomenon ubiquitous to single-emitters, is generally considered to be temporally random intensity fluctuations between "bright" ("On") and "dark" ("Off") states. However, individual quantum-dots (QDs) rarely exhibit such telegraphic signals, and yet, a vast majority of single-NC blinking data are analyzed using a single fixed threshold which generates binary trajectories. Furthermore, while blinking dynamics can vary dramatically over NCs in the ensemble, the extent of diversity in the exponents (mOn/Off) of single-particle On-/Off-time distributions (P(tOn/Off)), often used to validate mechanistic models of blinking, remains unclear due to a lack of statistically relevant data sets. Here, we subclassify an ensemble of QDs based on the emissivity of each emitter and subsequently compare the (sub)ensembles' behaviors. To achieve this, we analyzed a large number (>1000) of blinking trajectories for a model system, Mn+2 doped ZnCdS QDs, which exhibits diverse blinking dynamics. An intensity histogram dependent thresholding method allowed us to construct distributions of relevant blinking parameters (such as mOn/Off). Interestingly, we find that single QD P(tOn/Off)s follow either truncated power law or power law, and their relative proportion varies over subpopulations. Our results reveal a remarkable variation in mOn/Off amongst as well as within subensembles, which implies multiple blinking mechanisms being operational amongst various QDs. We further show that the mOn/Off obtained via cumulative single-particle P(tOn/Off) is distinct from the weighted mean value of all single-particle mOn/Off, evidence for the lack of ergodicity. Thus, investigation and analyses of a large number of QDs, albeit for a limited time span of a few decades, are crucial to characterize the spatial heterogeneity in possible blinking mechanisms.
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Here, we report the synthesis of a penta-selenopeptide consisting of five benzyl protected selenocysteine residues. This selenopeptide was well characterized by both one- and two-dimensional (D) NMR spectroscopies. We find that the solution conformation is enriched with ß-sheet structures, which have a propensity to self-assemble and form amyloid fibrils.
Subject(s)
Amyloid/chemistry , Peptides/chemistry , Selenocysteine/chemistry , Amyloid/chemical synthesis , Amyloid/ultrastructure , Chemistry Techniques, Synthetic , Circular Dichroism , Microscopy, Electron, Transmission , Peptides/chemical synthesis , Protein Structure, Secondary , Selenocysteine/chemical synthesis , Spectroscopy, Fourier Transform InfraredABSTRACT
Abrupt fluorescence intermittency or blinking is long recognized to be characteristic of single nano-emitters. Extended quantum-confined nanostructures also undergo spatially heterogeneous blinking; however, there is no such precedent in dimensionally unconfined (bulk) materials. Herein, we report multi-level blinking of entire individual organo-lead bromide perovskite microcrystals (volume=0.1-3â µm3 ) under ambient conditions. Extremely high spatiotemporal correlation (>0.9) in intracrystal emission intensity fluctuations signifies effective communication amongst photogenerated carriers at distal locations (up to ca. 4â µm) within each crystal. Fused polycrystalline grains also exhibit this intriguing phenomenon, which is rationalized by correlated and efficient migration of carriers to a few transient nonradiative traps, the nature and population of which determine blinking propensity. Observation of spatiotemporally correlated emission intermittency in bulk semiconductor crystals opens the possibility of designing novel devices involving long-range (mesoscopic) electronic communication.
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We use single-particle tracking to investigate colloidal dynamics in hybrid assemblies comprising colloids enmeshed in a crosslinked polymer network. These assemblies are prepared using ice templating and are macroporous monolithic structures. We investigate microstructure-property relations in assemblies that appear chemically identical but show qualitatively different mechanical response. Specifically, we contrast elastic assemblies that can recover from large compressive deformations with plastic assemblies that fail on being compressed. Particle tracking provides insights into the microstructural differences that underlie the different mechanical response of elastic and plastic assemblies. Since colloidal motions in these assemblies are sluggish, particle tracking is especially sensitive to imaging artifacts such as stage drift. We demonstrate that the use of wavelet transforms applied to trajectories of probe particles from fluorescence microscopy eliminates stage drift, allowing a spatial resolution of about 2 nm. In elastic and plastic scaffolds, probe particles are surrounded by other particles-thus, their motion is caged. We present mean square displacement and van Hove distributions for particle motions and demonstrate that plastic assemblies are characterized by significantly larger spatial heterogeneity when compared with the elastic sponges. In elastic assemblies, particle diffusivities are peaked around a mean value, whereas in plastic assemblies, there is a wide distribution of diffusivities with no clear peak. Both elastic and plastic assemblies show a frequency independent solid modulus from particle tracking microrheology. Here too, there is a much wider distribution of modulus values for plastic scaffolds as compared to elastic, in contrast to bulk rheological measurements where both assemblies exhibit a similar response. We interpret our results in terms of the spatial distribution of crosslinks in the polymer mesh in the colloidal assemblies.
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While fluorescence microscopy has become an essential tool amongst chemists and biologists for the detection of various analyte within cellular environments, non-uniform spatial distribution of sensors within cells often restricts extraction of reliable information on relative abundance of analytes in different subcellular regions. As an alternative to existing sensing methodologies such as ratiometric or FRET imaging, where relative proportion of analyte with respect to the sensor can be obtained within cells, we propose a methodology using spectrally-resolved fluorescence microscopy, via which both the relative abundance of sensor as well as their relative proportion with respect to the analyte can be simultaneously extracted for local subcellular regions. This method is exemplified using a BODIPY sensor, capable of detecting mercury ions within cellular environments, characterized by spectral blue-shift and concurrent enhancement of emission intensity. Spectral emission envelopes collected from sub-microscopic regions allowed us to compare the shift in transition energies as well as integrated emission intensities within various intracellular regions. Construction of a 2D scatter plot using spectral shifts and emission intensities, which depend on the relative amount of analyte with respect to sensor and the approximate local amounts of the probe, respectively, enabled qualitative extraction of relative abundance of analyte in various local regions within a single cell as well as amongst different cells. Although the comparisons remain semi-quantitative, this approach involving analysis of multiple spectral parameters opens up an alternative way to extract spatial distribution of analyte in heterogeneous systems. The proposed method would be especially relevant for fluorescent probes that undergo relatively nominal shift in transition energies compared to their emission bandwidths, which often restricts their usage for quantitative ratiometric imaging in cellular media due to strong cross-talk between energetically separated detection channels.
Subject(s)
Mercury/analysis , Boron Compounds , Cell Line, Tumor , Fluorescent Dyes , Humans , Microscopy, FluorescenceABSTRACT
While dynamics of single-molecule (SM) fluorescent probes have been used to investigate the structure and relaxation processes in polymers near the glass transition temperature (Tg), it is difficult to perform SM imaging at elevated temperatures which restricts such studies to a limited number of polymers for which Tg is close to room temperature (RT). Plasticization, solvent (or additive) induced lowering of Tg, offers an alternate avenue to access various effective temperatures in the glassy and rubbery phases of polymers under ambient conditions. By investigation of the reorientational propensity of individual Rhodamine 6G (Rh6G) probes, which is governed by rigidity/dynamics of the polymer cavities, we have explored the extent of spatiotemporal heterogeneity during moisture induced plasticization of poly(vinylpyrrolidone) (PVP), far below and near (below and above) bulk Tg. Lack of any probe reorientation suggests that the matrix remains extremely rigid up to a certain level of hydration, as expected for probes buried deep within the glassy state. At intermediate levels of hydration, SMs undergo a wide variety of rotational dynamics ranging from being static/wobbling motion to slow, hindered large-angle reorientation, as well as facile, intermittently hindered fast rotation, which reflects that swelling/softening of network cavities is spatiotemporally extremely diverse as the effective Tg approaches RT. SM probes exhibit temporally nonuniform rotational mobility even at relatively high moisture contents of the matrix beyond which probes can undergo translational motion, which indicates that relatively slow time scale polymer segmental motion can be operational for plasticized PVP (in the rubbery state). Our inferences are supported by the non-Gaussian nature of angular jump distributions for dipolar reorientation, similar to those reported for translational diffusion of SM tracers in polymers and cellular media, suggesting the existence of slow time-varying local environmental changes around individual probe molecules during plasticization.
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Water dispersible citrate-capped LaF3:Eu(5%) nanocrystals (NCs) have been partially surface-functionalized by 1,10-phenanthroline (phen) via a ligand exchange method to produce novel water dispersed citrate/phen-capped LaF3:Eu(5%) NCs in which citrate ligands preserve the water dispersibility of the NCs and phen ligands act as sensitizers of surface Eu(3+)-dopant sites. The partial ligand exchange and the formation of water dispersed NCs have been monitored by (1)H NMR spectroscopy, as well as luminescence measurements at different time intervals during the reaction. These NCs display a distinct phen-sensitized Eu(3+)-emission profile with enhanced intensity in water as compared to the emission profile and intensity obtained upon direct excitation. Time-resolved (or time-gated) emission spectroscopy (TRES) has been used to probe PL dynamics of Eu(3+)-sites of LaF3:Eu(5%) NCs by taking advantage of selectively sensitizing surface Eu(3+)-dopant sites by phen ligands as well as by exciting all the Eu(3+)-sites in the NCs upon direct excitation. TRES upon direct excitation of the citrate-capped LaF3:Eu(5%) NCs reveals that Eu(3+)-dopants occupy at least three different sites, each with a different emission profile and lifetime, and emission from purely interior Eu(3+)-sites has been resolved due to their long lifetime as compared to the lifetime of purely surface and near surface Eu(3+)-sites. In contrast, the phen-sensitized emission from citrate/phen-capped LaF3:Eu(5%) NCs displays similar emission profiles and lifetimes in TRES measurements, which reveal that phen truly sensitizes purely surface dopant sites of the NCs in water, all of which have nearly the same local environment. The phen-sensitized Eu(3+)-emission of the NCs in water remains stable even upon addition of various buffer solutions at physiological pH, as well as upon addition of water-miscible organic solvents. Furthermore, the two-photon excitation (λex. = 720 nm) of these water-soluble phen-capped NCs produces bright red Eu(3+) emission, which reveals that these NCs are promising for potential applications in biological imaging.