In vivo imaging with a fast large-area multiphoton exoscope (FLAME) captures the melanin distribution heterogeneity in human skin.

Melanin plays a significant role in the regulation of epidermal homeostasis and photoprotection of human skin. The assessment of its epidermal distribution and overall content is of great interest due to its involvement in a wide range of physiological and pathological skin processes. Among several spectroscopic and optical imaging methods that have been reported for non-invasive quantification of melanin in human skin, the approach based on the detection of two-photon excited fluorescence lifetime distinguishes itself by enabling selective detection of melanin with sub-cellular resolution, thus facilitating its quantification while also resolving its depth-profile. A key limitation of prior studies on the melanin assessment based on this approach is their inability to account for the skin heterogeneity due to the reduced field of view of the images, which results in high dispersion of the measurement values. Pigmentation in both normal and pathological human skin is highly heterogeneous and its macroscopic quantification is critical for reliable measurements of the epidermal melanin distribution and for capturing melanin-related sensitive dynamic changes as a response to treatment. In this work, we employ a fast large-area multiphoton exoscope (FLAME), recently developed by our group for clinical skin imaging, that has the ability to evaluate the 3D distribution of epidermal melanin content in vivo macroscopically (millimeter scale) with microscopic resolution (sub-micron) and rapid acquisition rates (minutes). We demonstrate significant enhancement in the reliability of the melanin density and distribution measurements across Fitzpatrick skin types I to V by capturing the intra-subject pigmentation heterogeneity enabled by the large volumetric sampling. We also demonstrate the potential of this approach to provide consistent measurement results when imaging the same skin area at different times. These advances are critical for clinical and research applications related to monitoring pigment modulation as a response to therapies against pigmentary skin disorders, skin aging, as well as skin cancers.

Stabilization of Mixed-Halide Lead Perovskites Under Light by Photothermal Effects.

Mixed-halide lead perovskites (MHLPs) are semiconductor materials with bandgaps that are tunable across the visible spectrum and have seen promising applications in photovoltaics and optoelectronics. However, their segregation into phases with enriched halide components, under resonant light illumination and/or electric field, have hindered their practical applications. Herein, we demonstrate the stabilization of the MHLP photoluminescence (PL) peak as a function of their excitation intensities. This effect is associated with the phase segregation of MHLPs and their subsequent remixing by photothermal heating. We conclude that the balance between these opposing processes dictates the equilibrium PL peak of the MHLPs. The findings in this work could serve as a potential approach to obtain MHLP with stable emission peaks under operating conditions.

Time-resolved temperature-jump measurements and steady-state thermal imaging of nanoscale heat transfer of gold nanostructures on AlGaN:Er3+ thin films.

For a nanostructure sitting on top of an AlGaN:Er3+ thin film, a new thermal imaging technique is presented where dual cameras collect bandpass filtered videos from the H and S bands of Er3+ emission. We combine this thermal imaging technique with our newly developed time-resolved temperature measurement technique which relies on luminescence thermometry using Er3+ emission. This technique collects time-resolved traces from the H and S bands of Er3+ emission. The H and S signal traces are then used to reconstruct the time-resolved temperature transient when a nanostructure is illuminated with a pulsed 532 nm light. Two different types of samples are interrogated with these techniques (drop-casted gold nanosphere cluster and lithographically prepared gold nanodot) on the AlGaN:Er3+ film. Steady-state and time-resolved temperature data are collected when the samples are immersed in air and water. The results of time-resolved temperature-jump measurements from a cluster of gold nanospheres show extremely slow heat transfer when the cluster is immersed in water and nearly 200-fold increase when immersed in air. The low thermal diffusivity for the cluster in water suggests poor thermal contact between the cluster and the thermal bath. The lithographically prepared nanodot has much better adhesion to the AlGaN film, resulting in much higher thermal diffusivity in both air and water. This proof-of-concept demonstration opens a new way to measure the dynamics of the local heat generation and dissipation at the nanoparticle-media interface.

Phase Segregation and Photothermal Remixing of Mixed-Halide Lead Perovskites.

Mixed-halide lead perovskites (MHPs) are promising materials for photovoltaics and optoelectronics due to their highly tunable band gaps. However, they phase segregate under continuous illumination or an electric field, the mechanism of which is still under debate. Herein we systematically measure the phase segregation behavior of polymer-encapsulated CH3NH3Pb(BrxI1-x)3 MHPs as a function of excitation intensity and the nominal halide ratio by in situ photoluminescence microspectroscopy and observe surprising phase dynamics at the beginning of the illumination. The initial phase segregation to I-rich and Br-rich phases is observed followed by the formation of a new mixed-halide phase within several seconds that has not been reported before. We propose that the photothermal effect is amplified at the small-size I-rich domains, which significantly changes the local phase segregation in the otherwise uniform film within milliseconds after illumination.

Cyanine-Based Polymer Dots with Long-Wavelength Excitation and Near-Infrared Fluorescence beyond 900 nm for In Vivo Biological Imaging.

Bioimaging in the near-infrared window is of great importance to study the dynamic processes in vivo with deep penetration, high spatiotemporal resolution, and minimal tissue absorption, scattering, and autofluorescence. In spite of the huge progress on the synthesis of small organic fluorophores and inorganic nanomaterials with emissions beyond 900 nm, it remains a tough challenge to synthesize semiconducting polymers with fluorescence over this region. Here, we synthesized a series of heptamethine cyanine-based polymers with both absorption and emission in the near-infrared region. We prepared these polymers as semiconducting polymer dots (Pdots) in pure water with great biocompatibility. The fluorescence quantum yield of the Pdots can be as high as 14% with a full width at half-maximum of 53 nm, and their single-particle brightness is more than 20 times higher than commercial quantum dots or ∼300 times brighter than Food and Drug Administration (FDA)-approved indocyanine green (ICG) dyes. We further demonstrated the use of cyanine-based Pdots for specific cellular labeling and long-term tumor targeting in mice. We anticipate that these cyanine-based ultrabright Pdots could open up an avenue for next generations of near-infrared fluorescent agents.

In Situ Sensing of Reactive Oxygen Species on Dye-Stained Single DNA Molecules under Illumination.

Reactive oxygen species (ROS) are a necessary evil in many biological systems and have been measured with fluorescent probes at the ensemble levels both in vitro and in vivo. Measuring ROS generated from a single molecule is important for mechanistic studies, yet measuring ROS near a dye-labeled single-molecule under illumination has been challenging. Here, we use CellROX, a group of ROS probes, to sense ROS near dye-stained DNA that has been flow-stretched and immobilized on a surface. ROS is responsible for the photodamage of DNA molecules under this circumstance. In this report, we confirmed the ROS sensing reaction in bulk solutions and optimized the conditions for single-molecule experiments including the selection of substrates, dye concentrations, probes in the CellROX series, excitation lasers, and emission filter-sets. We observed a correlation between ROS and the dye-labeled DNA and localized the ROS-activated CellROX probe molecules at both the ensemble level and the single-molecule level.

Single-Particle Organolead Halide Perovskite Photoluminescence as a Probe for Surface Reaction Kinetics.

Photoluminescence (PL) of organolead halide perovskites (OHPs) is sensitive to OHPs' surface conditions and is an effective way to report surface states. Literature has reported that at the ensemble level, the PL of photoexcited OHP nanorods declines under an inert nitrogen (N2) atmosphere and recovers under subsequent exposure to oxygen (O2). At the single-particle level, we observed that OHP nanorods photoblink at rates dependent on both the excitation intensity and the O2 concentration. Combining the two sets of information with the charge-trapping/detrapping mechanism, we are able to quantitatively evaluate the interaction between a single surface defect and a single O2 molecule using a new kinetic model. The model predicts that the photodarkening of OHP nanorods in the N2 atmosphere has a different mechanism than conventional PL quenching, which we call photo-knockout. This model provides fundamental insights into the interactions of molecular O2 with OHP materials and helps design a suitable OHP interface for a variety of applications in photovoltaics and optoelectronics.

Reusable Chemically-Micropatterned Substrates via Sequential Photoinitiated Thiol-Ene Reactions as Template for Perovskite Thin-Film Microarrays.

Patterning semiconducting materials are important for many applications such as microelectronics, displays, and photodetectors. Lead halide perovskites are an emerging class of semiconducting materials that can be patterned via solution-based methods. Here we report an all-benchtop patterning strategy by first generating a patterned surface with contrasting wettabilities to organic solvents that have been used in the perovskite precursor solution then spin-coating the solution onto the patterned surface. The precursor solution only stays in the area with higher affinity (wettability). We applied sequential sunlight-initiated thiol-ene reactions to functionalize (and pattern) both glass and conductive fluorine-doped tin oxide (FTO) transparent glass surfaces. The functionalized surfaces were measured with the solvent contact angles of water and different organic solvents and were further characterized by XPS, selective fluorescence staining, and selective DNA adsorption. By simply spin-coating and baking the perovskite precursor solution on the patterned substrates, we obtained perovskite thin-film microarrays. The spin-coated perovskite arrays were characterized by XRD, AFM, and SEM. We concluded that patterned substrate prepared via sequential sunlight-initiated thiol-ene click reactions is suitable to fabricate perovskite arrays via the benchtop process. In addition, the same patterned substrates can be reused several times until a favorable perovskite microarray is acquired. Among a few conditions we have tested, DMSO solvent and modified FTO surfaces with alternatively carboxylic acid and alkane is the best combination to obtain high-quality perovskite microarrays. The solvent contact angle of DMSO on carboxylic acid-modified FTO surface is nearly zero and 65±3° on octadecane modified FTO surface.

Kinetic Model under Light-Limited Condition for Photoinitiated Thiol-Ene Coupling Reactions.

Thiol-ene click chemistry has become a powerful paradigm in synthesis, materials science, and surface modification in the past decade. In the photoinitiated thiol-ene reaction, an induction period is often observed before the major change in its kinetic curve, for which a possible mechanism is proposed in this report. Briefly, light soaking generates radicals following the zeroth-order reaction kinetics. The radical is the reactant that initializes the chain reaction of thiol-ene coupling, which is a first-order reaction. Combining both and under the light-limited conditions, a surprising kinetics represented by a Gaussian-like model evolves that is different from the exponential model used to describe the first-order reaction of the final product. The experimental data are fitted well with the new model, and the reaction kinetic constants can be pulled out from the fitting.

The Role of Thermal Activation and Molecular Structure on the Reaction of Molecular Surfaces.

Though surface modifications of organic thin films dramatically improve optoelectronic device performance, chemistry at organic surfaces presents new challenges that are not seen in conventional inorganic surfaces. This work demonstrates that the subsurface of pentacene remains highly accessible, even to large adsorbates, and that three distinct reaction regimes (surface, subsurface, and bulk) are accessed within the narrow thermal range of 30-75 °C. Progression of this transition is quantitatively measured via polarization modulation infrared reflection absorption spectroscopy, and atomic force microscopy is used to measure the thin-film morphology. Together, they reveal the close relationship between the extent of the reaction and the morphology changes. Finally, the reaction kinetics of the pentacene thin film is measured with a series of adsorbates that have different reactivity and diffusivity in the thin film. The results suggest that reaction kinetics in the thin film is controlled by both the reactivity and the adsorbate diffusivity in the thin-film lattice, which is very different than the traditional solution kinetics that is dominated by the chemical activation barriers. Combined, these experiments guide efforts toward rationally functionalizing the surfaces of organic semiconductors to enable the next generation of flexible devices.