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Controlling the intensity of surface-enhanced Raman spectroscopy (SERS) in a systematic manner is the key to understanding, and even predicting, the dynamics of the imposing phenomenon. It has been...

Surface-enhanced Raman scattering (SERS) is a technique that can deliver label-free, real-time, and multiplex detection of target molecules. However, the development of this potential tool has been...

A small-sized chromophore, BTTA-2OH, manifesting favorable solubility, large two-photon excitation efficiency, and good fluorescence photostability was synthesized to label the membrane of living cells for visualizing the dynamic movement of membrane-related vesicles via a two-photon fluorescence imaging technique based on wavelength-tunable temporal-focusing multiphoton excitation microscopy.

Surface plasmon-coupled emission (SPCE) substrates to enhance the blinking fluorescence of spontaneously blinking fluorophores in single-molecule localization microscopy (SMLM) were fabricated to reduce the excitation power density requirement and reveal the distribution of fluorophore-labeled proteins on a plasma membrane with nanoscale-level resolution. The systemic investigation of the contribution of local field enhancement, modified quantum yield, and emission coupling yield through glass coverslip substrates coated with metal layers of different thicknesses revealed that the silver-layer substrate with a thickness of 44 nm produces the highest SPCE fluorescence in spontaneously blinking fluorophores, and it has a highly directional SPCE fluorescence, which helps improve the detection efficiency. Moreover, the uniform and surface-enhanced field created on the substrate surface is beneficial for fluorescence background reduction in single fluorophore detection and localization, as well as for revealing the real position of fluorophores. Consequently, compared with a glass coverslip substrate, the presented SPCE substrate demonstrated a fluorescence enhancement of 480% and an increase in blinking events from a single spontaneously blinking fluorophore; moreover, the required excitation power density for SMLM imaging was significantly reduced to 23 W cm-2 for visualizing the distribution of epidermal growth factor receptors (EGFRs) on the basal plasma membrane of A549 lung cancer cells with a localization precision of 19 ± 7 nm. Finally, the fluorophore-labeled EGFRs on the basal plasma membrane in the presence of PIKfyve-specific inhibitor treatment were explored using SPCE-SMLM imaging; the results revealed a distinct reduction in the density of localization events because of a decrease in EGFR abundance at the plasma membranes of the cells.

Reactive oxygen species (ROS)-induced oxidative stress leads to neuron damage and is involved in the pathogenesis of chronic inflammation in neurodegenerative diseases (NDs), such as Alzheimer's, Parkinson's, and amyotrophic lateral sclerosis. Researchers, therefore, are looking for antiinflammatory drugs and gene therapy approaches to slow down or even prevent neurological disorders. Combining therapeutics has shown a synergistic effect in the treatment of human diseases. Many nanocarriers could be designed for the simultaneous codelivery of drugs with genes to fight diseases. However, only a few researches have been performed in NDs. In this study, we developed a mesoporous silica nanoparticle (MSN)-based approach for neurodegenerative therapy. This MSN-based platform involved multiple designs in the targeted codelivery of (1) curcumin, a natural antioxidant product, to protect ROS-induced cell damage and (2) plasmid RhoG-DsRed, which is associated with the formation of lamellipodia and filopodia for promoting neurite outgrowth. At the same time, TAT peptide was introduced to the plasmid RhoG-DsRed via electrostatic interaction to elevate the efficiency of nonendocytic pathways and the nuclear plasmid delivery of RhoG-DsRed in cells for enhanced gene expression. Besides, such a plasmid RhoG-DsRed/TAT complex could work as a noncovalent gatekeeper. The release of curcumin inside the channel of the MSN could be triggered when the complex was dissociated from the MSN surface. Taken together, this MSN-based platform combining genetic and pharmacological manipulations of an actin cytoskeleton as well as oxidative stress provides an attractive way for ND therapy.

Temporal focusing multiphoton excitation microscopy (TFMPEM) is insufficient to perform efficient excitation of fluorophores with the different two-photon absorption spectra in the multi fluorophores imaging via an illuminated configuration of fixed wavelength and polarization direction of the pulse laser because of the a diffraction device. In this study, we overcome and causes it is not useful for in the practical application and its development. In order to implement the temporal focusing microscope system could be able to sever for the different excitation wavelength range of 700-1000 nm, we work on the optical design of the temporal focusing an integration stage and scanning mirror with automatic tuning system for any excitation wavelength. The novel TFMPEM has successfully provided dynamic spectral absorption, 3D two-photon or SHG images with the original ability of the high frame rate.

Transcription factor complex NF-κB (p65/p50) is localized to the cytoplasm by its inhibitor IκBα. Upon activation, the Rel proteins p65/p50 are released from IκBα and transported through nuclear pore to affect many gene expressions. While inhibitions of up or down stream signal pathways are often ineffective due to crosstalks and compensations, direct blocking of the Rel proteins p65/p50 has long been proposed as a potential target for cancer therapy. In this work, a nanoparticle/antibody complex targeting NF-κB is employed to catch the Rel protein p65 in perinuclear region and thus blocking the translocation near the nuclear pore gate. TAT peptide conjugated on mesoporous silica nanoparticles (MSN) help non-endocytosis cell-membrane transducing and converge toward perinuclear region, where the p65 specific antibody performed the targeting and catching against active NF-κB p65 effectively. The size of the p65 bound nanoparticle becomes too big to enter nucleus. Simultaneous treatment of mice with the hybrid MSN and doxorubicin conferred a significant therapeutic effect against 4T1 tumor-bearing mice. The new approach of anti-body therapy targeting on transcription factor with "nucleus focusing" and "size exclusion blocking" effects of the antibody-conjugated nanoparticle is general and may be applicable to modulating other transcription factors.

Dual-color two-photon excitation (TPE)-fluorescence imaging is used in conventional TFMPEM to observe specimens with different fluorophore labels. However, concerns have been raised about the excitation efficiency and selectivity of the fluorophores under fixed-wavelength excitation. This study presents a wavelength-switching approach using a scanning mirror, beam expander, and diffraction grating in the TFMPEM to switch the excitation wavelengths and match the optimal absorption of the fluorophores to acquire dynamic dual-color TPE-fluorescence images. The presented TFMPEM system was demonstrated to have an axial excitation confinement of 2.3–5.0 μm for excitation wavelengths of 730–1000 nm, and was used to visualize three-dimensional images of the vasculature of a mouse brain. The TPE efficiencies of different fluorophores were evaluated through TFMPEM imaging with excitation wavelength scanning to obtain their TPE spectra. Consequently, time-lapsed dual-color TFMPEM imaging was performed on rhodamine 6G (R6G)–poly(lactic-co-glycolic acid) (PLGA) nanoparticles and enhanced-yellow-fluorescent protein (EYFP)-tagged clathrin using excitation wavelengths at the maximum TPEs of R6G and EYFP, respectively. Our results revealed the PLGA-nanoparticle uptake of live cells via long-lived clathrin-coated plaques in clathrin-mediated endocytosis.

The desire to improve human lives has led to striking development in biosensing technologies. While the ongoing research efforts are mostly dedicated to enhancing speed and sensitivity of the sensor, a third consideration that has become increasingly important is compactness, which is strongly desired in emergency situations and personal health management. Surface plasmon resonance imaging (SPRi) is one of the few techniques that can potentially fulfill all the three goals, considering its multiplexed assay capability. However, miniaturizing SPRi biosensors remains elusive as it entails complicated optical gears. Here, we significantly slim the architecture of SPRi devices by visualizing the varied local density of states around analytes. The unusual detection scheme is realized by building a gain-assisted SPRi with InGaN quantum wells (QWs), where the QW-plasmon coupling efficiency hinges on localized refractive index variation. This new modality abolishes the prism, the polarizer, and the beam-tracking components in the most used Kretschmann configuration without compromising the performances.

Super-resolution imaging based on single-molecule localization microscopy combined with the surface plasmon polariton (SPP)-enhanced fluorescence of spontaneously blinking fluorophores was demonstrated to visualize the nanoscale-level positioning information of cell-adhesion-associated proteins. Glass substrates with a deposited silver layer were utilized to induce a SPP-enhanced field on the silver surface and significantly strengthen the fluorescence signals of the fluorophores by more than 300%. The illumination power density for localization imaging at a spatial resolution of 25 ± 11 nm was 31.6 W cm−2. This low illumination power density will facilitate the reduction of phototoxicity of the biospecimens for single-molecule localization imaging. The proposed strategy provides a uniform distribution of the SPP-enhanced field on the silver surface, enabling visualization of the spatial distribution of labeled proteins without interference caused by the enhanced field distribution.

Size distribution histograms of HSNs and HRP@HSNs. Calibration curve of fluorescence intensity versus RITC-HRP. Flow cytometry, cytotoxicity, and cell proliferation assays. Entrapment efficiency and loading capacity of HRP@HSNs. (DOC 450Â kb)

Continuous and simultaneous 3D single-particle movement and local pH detection in HeLa cells were demonstrated for the first time by combining fluorescent mesoporous silica nanoparticles (FMSNs) and a single-particle tracking (SPT) technique with a precision of ∼10 nm. FMSNs, synthesized by the co-condensation of both pH-sensitive and reference dyes with a silica/surfactant source, allow long-term reliable ratiometric pH measurements with a precision better than 0.3 pH unit because of their excellent brightness and stability. pH variation in the surrounding area of FMSNs during endocytosis was monitored in real-time. Acidification and low mobility of FMSNs were observed at the early endocytic stage, whereas basification and high mobility of FMSNs were observed at the late stage. Our results indicate that it is possible to monitor local pH changes in the environments surrounding nanoparticles during the cellular uptake process of FMSNs, which provides much needed information for designing an efficient drug delivery nanosystem.

RNA is a drug target involved in diverse cellular functions and viral processes. Molecules that inhibit the HIV TAR RNA-Tat protein interaction may attenuate Tat/TAR-dependent protein expression and potentially serve as anti-HIV therapeutics. By incorporating positively charged residues with mixed side chain lengths, we designed peptides that bind TAR RNA with enhanced intracellular activity. Tat-derived peptides that were individually substituted with positively charged residues with varying side chain lengths were evaluated for TAR RNA binding. Positively charged residues with different side chain lengths were incorporated at each Arg and Lys position in the Tat-derived peptide to enhance TAR RNA binding. The resulting peptides showed enhanced TAR RNA binding affinity, cellular uptake, nuclear localization, proteolytic resistance, and inhibition of intracellular Tat/TAR-dependent protein expression compared to the parent Tat-derived peptide with no cytotoxicity. Apparently, the enhanced inhibition of protein expression by these peptides was not determined by RNA binding affinity, but by proteolytic resistance. Despite the high TAR binding affinity, a higher binding specificity would be necessary for practical purposes. Importantly, altering the positively charged residue side chain length should be a viable strategy to generate potentially useful RNA-targeting bioactive molecules.

A three-dimensional (3D) single fluorescent particle tracking strategy based on temporal focusing multiphoton excitation microscopy (TFMPEM) combined with astigmatism imaging is proposed for delivering nanoscale-level axial information that reveals 3D trajectories of single fluorospheres in the axially-resolved multiphoton excitation volume without z-axis scanning. It provides the dynamical ability by measuring the diffusion coefficient of fluorospheres in glycerol solutions with a position standard deviation of 14 nm and 21 nm in the lateral and axial direction and a frame rate of 100 Hz. Moreover, the optical trapping force based on the TFMPEM is minimized to avoid the interference in the tracing measurements compared to that in the spatial focusing MPE approaches. Therefore, we presented a three dimensional single particle tracking strategy to overcome the limitation of the time resolution of the multiphoton imaging using fast frame rate of TFMPEM, and provide three dimensional locations of multiple particles using an astigmatism method.

The optical parameters of temporal-focusing multiphoton excitation microscopy (TFMPEM), which is capable of achieving varying wavelength excitation for multiple fluorophore measurement, was systematically examined to have good excitation performance. For this purpose, the approaches were adopted to quantitatively evaluate the grating groove density, focal length of the collimating lens and objective, and different excitation wavelengths. A grating with a groove density of 830 lines/mm enables the TFMPEM system to achieve a wavelength range of 700–1000 nm by adjusting the incident angle of the ultrafast laser on the grating; a diffraction efficiency of 81 ± 3% was obtained at this wavelength range. By using the 830 lines/mm grating, a collimating lens with a 500 mm focal length and a 60× water immersion objective, we achieved a large excitation area and a better filling effect of the spectrum band of the pulse on the back focal plane of the objective; these parameters ensure high optical sectioning and small variation in the illumination power density within this wavelength range. The variation ranges of the excitation area, optical sectioning, and illumination power density of 4272 ± 798 μm2, 2.6 ± 0.3 μm, and 71 ± 29%, respectively, were obtained in two-photon excitation fluorescence imaging at 700–1000 nm excitation wavelengths.

Herein, we describe a simple fabrication procedure for creating artificial hierarchical micro/nanopillars on silicon substrates that allows an effective, precise control of the interfacial adhesion and surface hydrophobicity. These well-defined hierarchical micro/nanostructures have four possible wetting states: Cassie-Cassie (C-C), Cassie-Wenzel (C-W), Wenzel-Cassie (W-C) and Wenzel-Wenzel (W-W). By controlling the critical height of the micro/nanopillars, it is possible to fabricate hierarchical micro/nanostructures in these four states. Thus, the hierarchical superhydrophobic surfaces proposed and fabricated in this study are promising for mimicking either lotus leaves with low adhesion or rose petals with high adhesion.

Wavelength tunable temporal focusing multiphoton excitation microscopy (TFMPEM) is conducted to visualize optical sectioning images of multiple fluorophore–labeled specimens through the optimal two-photon excitation (TPE) of each type of fluorophore. The tunable range of excitation wavelength was determined by the groove density of the grating, the diffraction angle, the focal length of lenses, and the shifting distance of the first lens in the beam expander. Based on a consideration of the trade-off between the tunable-wavelength range and axial resolution of temporal focusing multiphoton excitation imaging, the presented system demonstrated a tunable-wavelength range from 770 to 920 nm using a diffraction grating with groove density of 830 ?? lines / mm . TPE fluorescence imaging examination of a fluorescent thin film indicated that the width of the axial confined excitation was 3.0 ± 0.7 ?? ? m and the shifting distance of the temporal focal plane was less than 0.95 ?? ? m within the presented wavelength tunable range. Fast different wavelength excitation and three-dimensionally rendered imaging of Hela cell mitochondria and cytoskeletons and mouse muscle fibers were demonstrated. Significantly, the proposed system can improve the quality of two-color TFMPEM images through different excitation wavelengths to obtain higher-quality fluorescent signals in multiple-fluorophore measurements.

Flexible polymer nanopillar substrates were used to systematically demonstrate cell alignment and migration guided by the directional formation of focal adhesions. The polymer nanopillar substrates were constructed to various height specifications to provide an extensive variation of flexibility; a rectangular arrangement created spatial confinement between adjacent nanopillars, providing less spacing in the horizontal and vertical directions. Three polymer nanopillar substrates with the diameter of 400 nm and the heights of 400, 800, and 1200 nm were fabricated. Super-resolution localization imaging and protein pair–distance analysis of vinculin proteins revealed that Chinese hamster ovary (CHO) cells formed mature focal adhesions on 1200 nm high nanopillar substrates by bending adjacent nanopillars to link dot-like adhesions. The spacing confinement of the adjacent nanopillars enhanced the orthogonal directionality of the formation tendency of the mature focal adhesions. The directional formation of the mature focal adhesions also facilitated the organization of actin filaments in the horizontal and vertical directions. Moreover, 78% of the CHO cells were aligned in these two directions, in conformity with the flexibility and nanotopographical cues of the nanopillars. Biased cell migration was observed on the 1200 nm high nanopillar substrates.

Reactive oxygen species (ROS) are important factors in many clinical diseases. However, direct delivery of antioxidant enzymes into cells is difficult due to poor cell uptake. A proper design of delivery of enzymes by nanoparticles is very desirable for therapeutic purposes. To overcome the cell barrier problem, a designed mesoporous silica nanoparticle (MSN) system with attached TAT-fusion denatured enzyme for enhancing cell membrane penetration has been developed. Simultaneous delivery of two up-downstream antioxidant enzymes, superoxide dismutase (SOD) and glutathione peroxidase(GPx), reveals synergistic efficiency of ROS scavenging, compared to single antioxidant enzyme delivery. TAT peptide conjugation provided a facile nonendocytosis cell uptake and escape from endosome while moving and aggregating along the cytoskeleton that would allow them to be close to each other at the same time, resulting in the cellular antioxidation cascade reaction. The two-enzyme delivery shows a significant synergistic effect for protecting cells against ROS-induced cell damage and cell cycle arrest. The nanocarrier strategy for enzyme delivery demonstrates that intracellular anti-ROS cascade reactions could be regulated by multifunctional MSNs carrying image fluorophore and relevant antioxidation enzymes.

In the present study, we demonstrate the synthesis and applications of multifunctional gold nanorod-based probes for specific targeting and noninvasive imaging based on localized heating generated by gold nanorods after NIR irradiation. The structural design of the probe consists of MUA (11-mercaptoundecanoic acid)-capped gold nanorods covalently linked with low-molecular-weight chitosan oligosaccharide (M(w) ~5000) via carbodiimide (EDC) coupling agent. This surface modification is performed for complete replacement of toxic CTAB (hexadecyltrimethyl-ammonium chloride) and acid-responsive delivery of gold nanorods in acidic environment as known to be present at tumor surrounding areas. The resulting chitosan oligosaccharide-modified gold nanorods (CO-GNRs) were further conjugated with tumor targeting monoclonal antibody against EGFR (epidermal growth factor receptor) to provide localized targeting functionality owing to the overexpression of EGFR in human oral adenosquamous carcinoma cell line CAL 27. Initial in vitro and in vivo toxicity assessments indicated that CO-GNRs did not induce any significant toxicity and are thus suitable for biological applications. Furthermore, selective targeting and accumulation of CO-GNRs were observed in vitro via two-photon luminescence imaging studies in CAL 27, which was also observed through in vivo targeting studies performed via NIR (near-infrared) laser irradiation in CAL 27 xenografts of BALB/c nude mice. Hence, the CO-GNRs that we have developed are biocompatible and nontoxic and can be a potential candidate for in vivo targeted delivery, noninvasive imaging based on localized hyperthermia, and photothermal-related therapies.

In this study, we have explored how light trapping efficiency can be enhanced by using gold nanoparticles (Au NPs) of various sizes and shapes on the front of polymer solar cells (PSCs) with the active layer—blends of poly(3-hexyl thiophene) and [6,6]-phenyl-C61-butyric acid methyl ester. The light-concentrating behavior was enhanced after we had incorporated gold nanospheres or nanorods into the anodic buffer layer [based on poly(3,4-ethylenedioxythiophene):polystyrenesulfonate] to trigger various localized surface plasmon resonance (LSPR) bands. Comparison of the optical characteristics and the performance of the PSCs prepared with and without Au NPs, and we found that the UV–vis and wavelength-dependent photoluminescent spectral data corroborated with the device performance due to the photon management by considering the light scattering and LSPR effects at the active layer. The presence of Au NPs increased the power conversion efficiency to approximately 4.3% (an enhancement of 24%).

Mesoporous silica nanoparticles (MSNs) have received intensive recent interest as multi-functional nanocarriers for biological cells and tissues. Their cell uptake and intracellular distribution depends on the surface charge they carry. We develop a new approach for monitoring their uptake and intracellular localization by measuring the local pH value of the MSNs. We synthesized a ratiometric pH-sensor loaded with mesoporous silica nanoparticles that can map the location of the MSN through the site pH in HeLa cells. Positively charged MSN-TA was found to locate in higher pH regions which are identified as cytosol mostly. On the other hand, negatively charged MSN-PPs were trapped in acidic endosomes. Evidence is given for direct penetration of membranes through a charge-mediated membrane–nanoparticle interaction mechanism.

The formation of focal adhesions on various sizes of fibronectin patterns, ranging from 200 μm to 250 nm, was systematically investigated by total internal reflection fluorescence microscopy and super-resolution imaging. It was found that cells adhered to and spread on these micro/nanopatterns, forming focal adhesions. On a micrometer scale the shape of the focal adhesions was elongated. However, on the nanometer scale, the shape of focal adhesions became dotlike. To further explore the distribution of focal adhesion proteins formed on surfaces, a localization-based super-resolution imaging technique was employed in order to determine the position and density of vinculin proteins. A characteristic distance of 50 nm was found between vinculin molecules in the focal adhesions, which did not depend on the size of the fibronectin nanopatterns. This distance was found to be crucial for the formation of focal adhesions. In addition, the density of vinculin at the focal adhesions formed on the nanopatterns increased as the pattern size decreased. The density of the protein was found to be 425 ± 247, 584 ± 302, and 703 ± 305 proteins μm(-2) on the 600, 400, and 250 nm fibronectin patterns respectively. Whereas 226 ± 77 proteins μm(-2) was measured for the matured focal adhesions on homogeneous fibronectin coated substrates. The increase in vinculin density implies that an increase in mechanical load was applied to the focal adhesions formed on the smaller nanopatterns.

In this study, we used a transferring process to fabricate a simple light-harvesting system featuring 2D periodic granular-like electrodes for polymer solar cells (PSCs). This transferring technique, which was based on nanosphere lithography, could be used to fabricate periodic nanostructures on both the photoactive layers and the Al electrodes in the normal PSC device configuration (indium tin oxide glass/PEDOT:PSS/photoactive layer/Al). We investigated the properties of the PSC devices featuring periodic nanostructures in the photoactive layers using reflection UV–vis spectra and in terms of their external quantum efficiency (EQE) and photocurrent–voltage characteristics. In addition, we used numerical simulations to evaluate the electromagnetic field distributions in the devices. The light trapping efficiency in the PSCs featuring periodic nanostructures was enhanced as a result of light scattering and surface plasmon resonance effects. Relative to conventional devices featuring a flat geometry, the po...

Core/shell quantum dots (CdSe/Zns) conjugated with various nuclear localization signaling (NLS) peptides, which could facilitate the transportation of quantum dots across the plasma membrane into the nucleus, have been utilized to investigate the uptake mechanism of targeted delivery. Because of their brightness and photostability, it was possible to trace the trajectories of individual quantum dots in living cells using both confocal and total internal reflection microscopes. We found that, when the quantum dots were added to a cell culture, the peptide-coated quantum dots entered the cell nucleus while the uncoated quantum dots remained in the cytoplasm. At 8 nM, most of the peptide coated quantum dots were found in the cytoplasm due to aggregation. However, at a lower concentration (0.08 nM), approximately 25% of the NLS peptide-coated quantum dots entered the cell nucleus. We also found that some quantum dots without NLS coating could also enter the nucleus, suggesting that the size of the quantum dots may play an important role in such a process.

We have systematically explored how plasmonic effects influence the characteristics of polymer photovoltaic devices (OPVs) incorporating a blend of poly(3-hexylthiophene) (P3HT) and [6,6]-phenyl-C(61)-butyric acid methyl ester (PCBM). We blended gold nanoparticles (Au NPs) into the anodic buffer layer to trigger localized surface plasmon resonance (LSPR), which enhanced the performance of the OPVs without dramatically sacrificing their electrical properties. Steady state photoluminescence (PL) measurements revealed a significant increase in fluorescence intensity, which we attribute to the increased light absorption in P3HT induced by the LSPR. As a result, the rate of generation of excitons was enhanced significantly. Furthermore, dynamic PL measurements revealed that the LSPR notably reduced the lifetime of photogenerated excitons in the active blend, suggesting that interplay between the surface plasmons and excitons facilitated the charge transfer process. This phenomenon reduced the recombination level of geminate excitons and, thereby, increased the probability of exciton dissociation. Accordingly, both the photocurrents and fill factors of the OPV devices were enhanced significantly. The primary origin of this improved performance was local enhancement of the electromagnetic field surrounding the Au NPs. The power conversion efficiency of the OPV device incorporating the Au NPs improved to 4.24% from a value of 3.57% for the device fabricated without Au NPs.

We have synthesized a low-bandgap polymer, bithiazole-based polymer (PCPDTTBT) containing cyclopentadithiophene and thiophene units, and studied its opticophysical properties and morphologies in bulk heterojunction films after annealing. We used confocal optical microscopy in conjunction with a fluorescence module to record exciton lifetime images within photoactive layers of PCPDTTBT and [6,6]-phenyl-C61-butyric acid methyl ester (PCBM). These images, which were consistent with those recorded using atomic force microscopy and transmission electron microscopy, revealed that phase separation of PCPDTTBT and PCBM occurred during slow solvent evaporation. This phase separation not only provided bicontinuous pathways for carrier transport to the respective electrodes but also enhanced the degree of polymer chain stacking, thereby improving the absorption and balancing the electron and hole mobilities. We tested the performance of solar cells incorporating PCPDTTBT and various fullerene derivatives. Under AM 1...

In this study, the exciton lifetime images within the photoactive layers of poly(3-hexylthiophene) (P3HT) and [6,6]-phenyl-C61-butyric acid methyl ester (PCBM) are revealed by confocal optical microscopy combined with the fluorescence module. The images reveal that the active layers during slow solvent evaporation provide 3D pathways for charge transport and the concentration gradient through the film which reflects the better cell performance. This technique offers a great help to investigate the 3D optical-physical property without destroying the blends.

Control of blend morphology at the nanoscale and high charge mobility is essential for polymer photovoltaic devices in terms of their power conversion efficiencies (PCE). In the case of bulk heterojunctions solar cells, both blend morphology and charge mobility are influenced by thermal treatment. In this manuscript, we study the effects of annealing temperature on polymer PV devices with blends of poly[9,9’- dioctyl-fluorene-co-bithiophene] (F8T2) and [6,6]-phenyl-C 61 -butyric acid methyl ester (PCBM). The morphological changes of blended films were observed upon thermal annealing temperature near and above glass transition temperature (130 °C). Such microstructural transformations resulted in modified charge transport pathways and therefore greately influenced the device performance. The highest PCE of 2.14% with an open-circuit voltage ( V OC ) of 0.99 V and a short-circuit current ( J SC ) of 4.24 mA/cm 2 was achieved by device annealing at 70 °C for 20 min.

Replication forks are vulnerable to wayward nuclease activities. We report here our discovery of a new member in guarding genome stability at replication forks. We previously isolated a Drosophila mutation, wuho (wh, no progeny), characterized by a severe fertility defect and affecting expression of a protein (WH) in a family of conserved proteins with multiple WD40 repeats. Knockdown of WH by siRNA in Drosophila, mouse, and human cultured cells results in DNA damage with strand breaks and apoptosis through ATM/Chk2/p53 signaling pathway. Mice with mWh knockout are early embryonic lethal and display DNA damage. We identify that the flap endonuclease 1 (FEN1) is one of the interacting proteins. Fluorescence microscopy showed the localization of WH at the site of nascent DNA synthesis along with other replication proteins, including FEN1 and PCNA. We show that WH is able to modulate FEN1’s endonucleolytic activities depending on the substrate DNA structure. The stimulatory or inhibitory effects of WH on FEN1’s flap versus gap endonuclease activities are consistent with the proposed WH’s functions in protecting the integrity of replication fork. These results suggest that wh is a new member of the guardians of genome stability because it regulates FEN1’s potential DNA cleavage threat near the site of replication., The essential protein Wuho can interact with FEN1 and switch its activity from potentially damaging gap endonuclease to useful flap endonuclease, thereby helping to protect the integrity of the replication fork and maintain genome stability., Author Summary Accurate genome replication is essential for the transmission of genetic information, and this process is vulnerable to factors that can induce DNA damage or block replication. To avoid this and achieve genome stability, many enzymes need to coordinate their activities at the replication forks (the area where the two DNA strands separate to replicate), thereby ensuring high efficiency and fidelity. Replication occurs simultaneously in both strands, but DNA polymerases only work in one direction (5′-to-3′), and while synthesis of one strand proceeds continuously, synthesis of the other must occur via small DNA fragments that need to be joined together after the removal of their 5′ ends by the flap endonuclease 1 (FEN1). In addition, FEN1 has a gap endonuclease activity that can potentially introduce strand breaks at the sites of replication. Here we report the identification of a new member of this complex of proteins called Wuho (WH) that has a role in guarding the genome stability at the replication forks. Down-regulation of Wuho in tissue culture cells results in double-strand DNA breaks and programmed cell death. Genetic and biochemical results indicate that WH interacts with FEN1 and regulates its activities at the replication fork by promoting its flap endonuclease activity while dampening its gap endonuclease activity. These results suggest a critical role of Wuho in protecting the integrity of replication forks.

This study develops a coupled waveguide-surface plasmon resonance (CWSPR) biosensor with a subwavelength grating structure for the real-time analysis of biomolecular interactions. In the proposed optical metrology system, normally incident white light is coupled into the waveguide layer through the subwavelength grating structure thereby enhancing the wave vector which excites the surface plasmons on the metal sensing surface. The proposed CWSPR biosensor not only retains the same sensing sensitivity as that of a conventional surface plasmon resonance device, but also yields a sharper dip in the reflectivity spectrum and therefore provides an improved measurement precision. Moreover, the metrology setup overcomes the limitations of the conventional Kretschmann attenuated total reflection approach and is less sensitive to slight variations in the angle of the incident light. The experimental results confirm that the current CWSPR biosensor provides a straightforward yet powerful technique for real-time biomolecular interaction analysis.

Surface thiolation affects the size of gold nanoparticles and the presence of visible luminescence under UV stimulation. We explored these phenomena by analysing alkanethiolate coatings with different carbon chain lengths, from 3-mercaptopropionic acid to 16-mercaptohexadecanoic acid, synthesized by intense X-ray irradiation. Photoluminescence is present for the smallest nanoparticles, but its intensity becomes more intense as the carbon chain length increases, achieving a quantum efficiency of 28% with a 16-mercaptohexadecanoic acid coating.

This paper presents a novel microfluidic system with integrated molecular imprinting polymer (MIP) films designed for surface plasmon resonance (SPR) biosensing of multiple nanoscale biomolecules. The innovative microfluidic chip uses pneumatic microvalves and micropumps to transport a precise amount of the biosample through multiple microchannels to sensing regions containing the locally spin-coated MIP films. The signals of SPR biosensing are basically proportional to the number of molecules adsorbed on the MIP films. Hence, a precise control of flow rates inside microchannels is important to determine the adsorption amount of the molecules in the SPR/MIP chips. The integration of micropumps and microvalves can automate the sample introduction process and precisely control the amount of the sample injection to the microfluidic system. The proposed biochip enables the label-free biosensing of biomolecules in an automatic format, and provides a highly sensitive, highly specific and high-throughput detection performance. Three samples, i.e. progesterone, cholesterol and testosterone, are successfully detected using the developed system. The experimental results show that the proposed SPR/MIP microfluidic chip provides a comparable sensitivity to that of large-scale SPR techniques, but with reduced sample consumption and an automatic format. As such, the developed biochip has significant potential for a wide variety of nanoscale biosensing applications.

This study utilizes a surface plasmon resonance (SPR) biosensing and a theoretical secondary structure calculation to investigate the influence of secondary structures on the DNA hybridization. It is found that the SPR angular shifts associated with the three pairs of 60mer oligonucleotides with prominent secondary structures are lower than those observed for the two pairs of oligonucleotides with no obvious secondary structures. It is also determined that increasing the DNA hybridization temperature from 35 to 45 °C reduces secondary structure effects. On the hybridization with mixture target oligonucleotides, the SPR results demonstrate that secondary structures interfere significantly.

Cyanine dye J-aggregate films are a class of absorbing and luminescent materials which have been extensively applied in the polariton-based research. Here we systematically study the DEDOC cyanine dyes J-aggregate films made by layer-by-layer assembly and spin-coating processes to establish a clear correlation between the film structure and the absorption and luminescence properties. From detailed analyses of morphology, optical spectra, and light-emitting diode characteristics, we demonstrate that layer-by-layer assembled film has higher degrees of homogeneity and molecular packing quality than spin-coated film, leading to a higher absorption coefficient, more uniform luminescence, and a greater electroluminescence quantum efficiency with maximized thickness.

A three-dimensional (3D) single fluorescent particle tracking strategy based on temporal focusing multiphoton excitation microscopy (TFMPEM) combined with astigmatism imaging is proposed for delivering nanoscale-level axial information that reveals 3D trajectories of single fluorospheres in the axially-resolved multiphoton excitation volume without z-axis scanning. Whereas other scanning spatial focusing multiphoton excitation schemes induce optical trapping interference, temporal focusing multiphoton excitation produces widefield illumination with minimum optical trapping force on the fluorospheres. Currently, the lateral and axial positioning resolutions of the dynamic particle tracking approach are about 14 nm and 21 nm in standard deviation, respectively. Furthermore, the motion behavior and diffusion coefficients of fluorospheres in glycerol solutions with different concentrations are dynamically measured at a frame rate up to 100 Hz. This TFMPEM with astigmatism imaging holds great promise for exploring dynamic molecular behavior deep inside biotissues via its superior penetration, reduced trapping effect, fast frame rate, and nanoscale-level positioning.

Galectin-3 has been reported to regulate the functions of a number of immune cell types. We previously reported that galectin-3 is translocated to immunological synapses in T cells upon T-cell receptor engagement, where it associates with ALG-2-interacting protein X (Alix). Alix is known to coordinate with the endosomal sorting complex required for transport (ESCRT) to promote human immunodeficiency virus (HIV)-1 virion release. We hypothesized that galectin-3 plays a role in HIV-1 viral budding. Cotransfection of cells of the Jurkat T line with galectin-3 and HIV-1 plasmids resulted in increased HIV-1 budding, and suppression of galectin-3 expression by RNAi in Hut78 and primary CD4+ T cells led to reduced HIV-1 budding. We used immunofluorescence microscopy to observe the partial colocalization of galectin-3, Alix and Gag in HIV-1-infected cells. Results from co-immunoprecipitation experiments indicate that galectin-3 expression promotes Alix-Gag p6 association, whereas the results of Alix knockdown suggest that galectin-3 promotes HIV-1 budding through Alix. HIV-1 particles released from galectin-3-expressing cells acquire the galectin-3 protein in an Alix-dependent manner, with proteins primarily residing inside the virions. We also found that the galectin-3 N-terminal domain interacts with the proline-rich region of Alix. Collectively, these results suggest that endogenous galectin-3 facilitates HIV-1 budding by promoting the Alix-Gag p6 association.

It is well known that ligand binding to the high-affinity GM-CSF receptor (GMR) activates JAK2. However, how and where this event occurs in a cellular environment remains unclear. Here, we demonstrate that clathrin- but not lipid raft-mediated endocytosis is crucial for GMR signaling. Knockdown expression of clathrin heavy chain or intersectin 2 (ITSN2) attenuated GMR-mediated activation of JAK2, whereas inhibiting clathrin-coated pits or plagues to bud off the membrane by the dominant-negative mutant of dynamin enhanced such event. Moreover, unlike the wild-type receptor, an ITSN2-non-binding mutant of GMR defective in targeting to clathrin-coated pits or plagues [collectively referred to as clathrin-coated structures (CCSs) here] failed to activate JAK2 at such locations. Additional experiments demonstrate that ligand treatment not only enhanced JAK2/GMR association at CCSs, but also induced a conformational change of JAK2 which is required for JAK2 to be activated by CCS-localized CK2. Interestingly, ligand-independent activation of the oncogenic mutant of JAK2 (JAK2V617F) also requires the targeting of this mutant to CCSs. But JAK2V617F seems to be constitutively in an open conformation for CK2 activation. Together, this study reveals a novel functional role of CCSs in GMR signaling and the oncogenesis of JAK2V617F.

Here, we report the facile preparation of tunable magnetic Ni-doped near-infrared (NIR) quantum dots (MNIR-QDs) as an efficient probe for targeting, imaging, and cellular sorting applications. We synthesized the MNIR-QDs via a hot colloidal synthesis approach to yield monodisperse and tunable QDs. These hydrophobic QDs were structurally and compositionally characterized and further functionalized with amino-PEG and carboxyl-PEG to improve their biocompatibility. Since QDs are known to be toxic due to the presence of cadmium, we have evaluated the in vitro and in vivo toxicity of our surface-functionalized MNIR-QDs. Our results revealed that surface-functionalized MNIR-QDs did not exhibit significant toxicity at the concentrations used in the experiments and are therefore suitable for biological applications. For further in vitro applications, we covalently linked folic acid to the surface of amino-PEG-coated MNIR-QDs through NHS chemistry to target the folate receptors largely present in the HeLa cells to demonstrate the specific targeting and magnetic behavior of these MNIR-QDs. Improved specificity has been observed with treatment of HeLa cells with the folic acid-linked amino PEG-coated MNIR QDs (FA-PEG-MNIR-QDs) compared to the one without folic acid. Since the synthesized probe has magnetic property, we have also successfully demonstrated sorting between the cells which have taken up the probe with the use of a magnet. Our findings strongly suggest that these functionalized MNIR-QDs can be a potential probe for targeting, cellular sorting, and bioimaging applications.

This paper reports the development of a novel force measurement device based on polymeric nanopillar arrays. The device was fabricated by a process combining nanosphere lithography, oxygen plasma treatment, deep etching and nano-molding. Well-ordered polymeric nanopillar arrays with various diameters and aspect ratios were fabricated and used as cell culture substrates. Cell traction forces were measured by the deflection of the nanopillars. Since the location of the nanopillars can be monitored at all times, this device allows for the measurement of the evolution of adhesion forces over time.

An energy upconversion system based on triplet–triplet annihilation exploiting an organic triplet sensitizer is devised and has achieved a white-light emission with a low power laser excitation.

We have developed a nanoparticle based Raman probe (Ag@Nile Red), which was capable of targeting the lipids, therefore, revealing the location of lipid droplets in live Caenorhabditis elegans (C. elegans).

The blinking effects of quantum dots have been utilized to localize individual quantum dots, which couldn't be resolved by conventional microscopy. It has been demonstrated that quantum dots could be localized with sub-10 nanometer resolution.

We report on red phosphorescent trilayer organic light-emitting field-effect transistors (LEFETs), which exhibit ambipolar transport characteristics but strong emission in the unipolar hole regime, with an exceptionally wide recombination zone of 60–70 µm and a maximum external quantum efficiency of 0.21%. From the results of detailed electroluminescence characterization, we clarify how the energy-level matching condition and transport geometry of the heterostructure govern the charge distribution and recombination, and affect the overall device performance.