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Cadmium-free quantum-dot (QD) fluorophores can bridge the gap between the macroscopic and microscopic domains in fluorescence super-resolution bioimaging. InP/ZnSe/ZnS QD photoblinking fluorescent probes can improve the performance of reactive super-resolution imaging techniques and spontaneously switch fluorophores between at least two states (open and close) without depending on intense laser light and specialized buffers for bioimaging. Multifocal structured illumination microscopy (MSIM) provides a two-fold resolution enhancement in sub-diffraction imaging, but higher resolutions are limited by the pattern frequency and signal-to-noise ratio. We exploit the synergy between MSIM and spontaneously switching InP/ZnSe/ZnS QD fluorophores to further increase the imaging resolution. We demonstrate the experimental combination of optical-fluctuation-enhanced super-resolution MSIM using ultrasonic-oscillation-assisted organic solvothermal synthesis of narrowband photoblinking InP/ZnSe/ZnS QDs. The InP/ZnSe/ZnS QDs show a monodisperse grain size of approximately 9 nm, fluorescence quantum yields close to 100%, and full width at half maximum below 30 nm. The structural, electronic, and optical properties are characterized through experiments and first-principles calculations. The enhanced MSIM imaging achieves an approximate fourfold improvement in resolution for fixed cells compared with widefield imaging. The proposed InP/ZnSe/ZnS QD fluorescent probes seem promising for super-resolution imaging using MSIM.

Photodynamic therapy (PDT) has been widely used in preclinical trials for treating various tumors. However, the hypoxic environment of tumors and the limited penetration depth of ultraviolet light severely weaken the PDT effect. To solve the above problems, a near-infrared (NIR) light-triggered oxygen (O

Photodynamic therapy (PDT) for hypoxic tumors has attracted wide attention owing to its noninvasiveness, easy maneuverability, and instantaneity. However, hypoxia in tumors and penetration depth of conventional ultraviolet light has greatly weakened its performance. To solve these problems, under NIR-II light irradiation, squaric acid nanoparticles (SQ NPs) with superior reactive oxygen, especially, hydroxyl radicals (•OH) production performance were first utilized for hypoxic tumor therapy. SQ NPs with intense light capture capability, intense NIR emission, and excellent photobleaching resistance show continuous •OH generation capabilities under NIR-II laser excitation. Through the superior PDT performance, the growth of hypoxic tumors was effectively inhibited, and the survival rate of mice was improved. This work highlights the application of NIR-II photoexcitation in deep tissue type I photodynamic therapy of hypoxic tumors, which will facilitate the development of hypoxic tumor PDT in deep depth.

Conjugated polymers with narrow band gaps are particularly useful for sorting and discriminating semiconducting single-walled carbon nanotubes (s-SWCNT) due to the low charge carrier injection barrier for transport. In this paper, we report two newly synthesized narrow-band-gap conjugated polymers (PNDITEG-TVT and PNDIC8TEG-TVT) based on naphthalene diimide (NDI) and thienylennevinylene (TVT) building blocks, decorated with different polar side chains that can be used for dispersing and discriminating s-SWCNT. Compared with the mid-band-gap conjugated polymer PNDITEG-AH, which is composed of naphthalene diimide (NDI) and head-to-head bithiophene building blocks, the addition of a vinylene linker eliminates the steric congestion present in head-to-head bithiophene, which promotes backbone planarity, extending the π-conjugation length and narrowing the band gap. Cyclic voltammetry (CV) and density functional theory (DFT) calculations suggest that inserting a vinylene group in a head-to-head bithiophene efficiently lifts the highest occupied molecular orbital (HOMO) level (-5.60 eV for PNDITEG-AH, -5.02 eV for PNDITEG-TVT, and -5.09 eV for PNDIC8TEG-TVT). All three polymers are able to select for s-SWCNT, as evidenced by the sharp transitions in the absorption spectra. Field-effect transistors (FETs) fabricated with the polymer:SWCNT inks display p-dominant properties, with higher hole mobilities when using the NDI-TVT polymers as compared with PNDITEG-AH (0.6 cm2V-1s-1for HiPCO:PNDITEG-AH, 1.5 cm2V-1s-1for HiPCO:PNDITEG-TVT, and 2.3 cm2V-1s-1for HiPCO:PNDIC8TEG-TVT). This improvement is due to the better alignment of the HOMO level of PNDITEG-TVT and PNDIC8TEG-TVT with that of the dominant SWCNT specie.

Optical fibers are typically used in telecommunications services for transmission of data, fiber tags are essential to distinguish between different transmission fibers or channels to ensure the working functions of the communication system. Traditional physical entity marking method for fiber labelling is bulk in size, easy confusion and most importantly the label information can be easilly accessed by all potential users. In this work, we propose an encrypted optical fiber tag based on encoded fiber Bragg grating array fabricated by a pointby-point femtosecond laser pulse chain inscription method. Gratings with different resonant wavelengths and reflectivities can be achieved by adjusting the grating period and refractive index modulations. It is demonstrated a binary data sequence carried by a fiber tag can be inscribed into the fiber core in the form of FBG array, and the data can be encryted by proper design of the spatial distributions of FBGs with varing reflection wavelengths and reflectivities.The proposed fiber tag technology could be used for port identification, encrypted data storage and transmission in fiber networks.

Structured illumination microscopy (SIM) is suitable for biological samples because of its relatively low-peak illumination intensity requirement and high imaging speed. The system resolution is affected by two typical detection modes: Point detection and area detection. However, a systematic analysis of the imaging performance of the different detection modes of the system has rarely been conducted. In this study, we compared laser point scanning point detection (PS-PD) and point scanning area detection (PS-AD) imaging in nonconfocal microscopy through theoretical analysis and simulated imaging. The results revealed that the imaging resolutions of PS-PD and PS-AD depend on excitation and emission point spread functions (PSFs), respectively. Especially, we combined the second harmonic generation (SHG) of point detection (P-SHG) and area detection (A-SHG) with SIM to realize a nonlinear SIM-imaging technique that improves the imaging resolution. Moreover, we analytically and experimentally compared the nonlinear SIM performance of P-SHG with that of A-SHG.

There is an urgent need for developing rapid and affordable antibiotic susceptibility testing (AST) technologies to inhibit the overuse of antibiotics. In this study, a novel microcantilever nanomechanical biosensor based on Fabry–Pérot interference demodulation was developed for AST. To construct the biosensor, a cantilever was integrated with the single mode fiber in order to form the Fabry–Pérot interferometer (FPI). After the attachment of bacteria on the cantilever, the fluctuations of cantilever caused by the bacterial movements were detected by monitoring the changes of resonance wavelength in the interference spectrum. We applied this methodology to Escherichia coli and Staphylococcus aureus, showing the amplitude of cantilever’s fluctuations was positively related on the quantity of bacteria immobilized on the cantilever and associated with the bacterial metabolism. The response of bacteria to antibiotics was dependent on the types of bacteria, the types and concentrations of antibiotics. Moreover, the minimum inhibitory and bactericidal concentrations for Escherichia coli were obtained within 30 minutes, demonstrating the capacity of this method for rapid AST. Benefiting from the simplicity and portability of the optical fiber FPI-based nanomotion detection device, the developed nanomechanical biosensor in this study provides a promising technique for AST and a more rapid alternative for clinical laboratories.

Rational design of tumor-microenvironment (TME)-activated nanoformulation for precisely targeted cancer treatment has recently attracted an enormous attention. However, the all-in-one TME-activated theranostic nanosystems with a simple preparation and high biocompatibility are still rarely reported. Herein, catalase nanocrystals (CatCry) are first introduced as a tumor microenvironment activatable nanoplatform for selective theranostics of colon cancer. They are engaged as (i) a "nanoreactor" for silver nanoparticles (AgNP) synthesis, (ii) a nanovehicle for tumor delivery of anticancer drug doxorubicin (DOX), and (iii) an

Research on dynamic events in living cells, such as intracellular transportation, is important for understanding cell functions. As movements occur within cells, the microenvironment of the moving vesicles or biomacromolecules may affect the behavior of them. Herein, we propose a method of simultaneously monitoring changes in spatial positions and the local environment related to the fluorescence lifetime, i.e., four-dimensional (4D) multi-particle parallel-tracking in living cells. Based on double-helix point spread function (DH-PSF) microscopy and streak camera, the method combines three-dimensional (3D) localization methods and fluorescence lifetime imaging. By modifying the PSF of the system, the 3D positions and fluorescence lifetime information for several molecules within a depth of a few microns can be acquired simultaneously from a single snapshot. The feasibility of this method is verified by simulating the real-time tracking of a single particle with a given trajectory. In addition, a proof-of-concept 4D tracking system based on the DH-PSF and streak camera was built. The experimental results show that the 3D localization and lifetime precision are σ(x, y, z) = (26 nm, 35 nm, 53 nm) and σ(τ) = 103 ps, respectively, and the effective depth of field is approximately 4 μm. Finally, intracellular endocytosis in a living cell was observed using the system, which demonstrated the successful 4D tracking of two microspheres moving within an axial depth of 4 μm. This work opens a new perspective for research of dynamic processes, by providing information about the chemical (microenvironments) and physical (positions) changes of moving targets in living cells.

Lipid droplets (LDs), a dynamic organelle, are of vital importance in regulating the storage of neutral lipids and energy homeostasis. The aberrant expression of LDs is found to be highly associated with diverse metabolic diseases. Thus, detecting and monitoring LDs are essential to study the pathological and physiological processes of LDs in living bodies. However, it remains challenging to obtain suitable imaging probes to track LDs in vivo. Fortunately, the emergence of carbon dots (CDs), which are fluorescent nanomaterials with good biocompatibility and high stability, has provided us an unprecedented choice. In this work, CDs were synthesized via a solvothermal treatment of commercial reagents, 3-dimethylaminophenol. Interestingly, the prepared CDs show an intense red emission in non-hydrogen-bonding solution and have strong LD-targeting ability without any postmodification of ligands. Moreover, due to their low phototoxicity and excellent photostability, CDs were successfully applied to track the dynamics of LDs in live cells and image LDs in different cell lines and lipid-rich tissues. Overall, this work here proposed an LD-specific red-emitting CD probe, which will be of great value for learning more about LD-associated behaviors and diseases.

This paper describes the effectiveness of donor-acceptor (D-A) conjugated polymers to disperse and select for semiconducting single-walled carbon nanotubes (s-SWCNTs) when enhanced by the inclusion of polar oligoethylene glycol-based side chains, without altering the D-A backbone. We designed and synthesized two sets of naphthalenediimide(NDI)-alt-bithiophene(T2)-based conjugated polymers with one of two alkyl side chains (decyl and dodecyl chains) of different lengths and with or without polar triethylene glycol side chains. The resulting low-band-gap copolymers all effectively disperse and select for s-SWCNT, but the inclusion of polar side chains enhances the interactions between the polymer backbone and the walls of the s-SWCNTs relative to the polymers with only alkyl side chains. As a result, the wrapping and selection efficiency of the polymer-SWCNT systems with polar side chains are both significantly enhanced. We further optimized the binding energy and surface coverage by combining glycol ether and dodecyl side chains to maximize wrapping efficiency, leading to a field-effect mobility of 2.82 cm2 V-1 s-1 and on/off current ratios of ∼2 × 107 in polymer-wrapped SWCNTs. Our results provide insight into the role of the side-chain interactions in the polymer wrapping and dispersion technique, and, because we focus on manipulating side chains, they can be generalized for other conjugated polymer backbones.

NaYF4 and LiYF4 nanoparticles co-doped with Pr3+ and Yb3+ are reported, manifesting 980 nm excited downconversion luminescence peaked at 1320 nm for NIR-II bioimaging and 447 nm excited upconversion luminescence at 275 nm for germicide action.

Hepatocellular carcinoma (HCC) is the most common primary liver cancer, which is not sensitive to radiotherapy and chemotherapy and very often experiences postoperative relapse. In this regard, effective screening of liver cancer is considered as the most important and urgent task. The aim of our study was to determine whether N-methyl-D-aspartate receptor (NMDAR) and, in particular, its subunits, can serve as biomarkers to distinguish the precancerous liver at early stages of liver fibrosis. We assessed the development of HCC after 10, 15 and 22 weeks using a HCC rat model. The expression of NMDAR subunits was monitored at different stages of HCC by means of immunohistochemistry combined with epifluorescence microscopy imaging, Western blotting and direct bisulfite sequencing. NMDAR subunits were not found in healthy liver tissues. In contrast, NMDAR subunits, in particular NR1 and NR2B, appeared at the stage of severe liver fibrosis (precancerous liver disease) in rats and were expressed during the development of HCC in rats and mice. Using the direct bisulfite sequencing, we detected that increased expression of NMDAR directly correlated with the demethylation of CpG islands in the promoter region of genes encoding receptor subunits. The obtained results confirmed that NMDAR subunits can serve as new biomarkers of precancerous liver disease, severe fibrosis, and its progression towards HCC.

Tunneling nanotubes (TNTs) are nanoscale, actin-rich, transient intercellular tubes for cell-to-cell communication, which transport various cargoes between distant cells. The structural complexity and spatial organization of the involved components of TNTs remain unknown. In this work, the STORM super-resolution imaging technique was applied to elucidate the structural organization of microfilaments and microtubules in intercellular TNTs at the nanometer scale. Our results reveal different distributions of microfilaments and intertwined structures of microtubules in TNTs, which promote the knowledge of TNT communications.

Fourier light-field microscopy (FLFM) uses a microlens array (MLA) to segment the Fourier plane of the microscopic objective lens to generate multiple two-dimensional perspective views, thereby reconstructing the three-dimensional (3D) structure of the sample using 3D deconvolution calculation without scanning. However, the resolution of FLFM is still limited by diffraction, and furthermore, it is dependent on the aperture division. In order to improve its resolution, a super-resolution optical fluctuation Fourier light-field microscopy (SOFFLFM) was proposed here, in which the super-resolution optical fluctuation imaging (SOFI) with the ability of super-resolution was introduced into FLFM. SOFFLFM uses higher-order cumulants statistical analysis on an image sequence collected by FLFM, and then carries out 3D deconvolution calculation to reconstruct the 3D structure of the sample. The theoretical basis of SOFFLFM on improving resolution was explained and then verified with the simulations. Simulation results demonstrated that SOFFLFM improved the lateral and axial resolution by more than [Formula: see text] and 2 times in the second- and fourth-order accumulations, compared with that of FLFM.

Super-resolution optical imaging is crucial to the study of cellular processes. Current super-resolution fluorescence microscopy is restricted by the need of special fluorophores or sophisticated optical systems, or long acquisition and computational times. In this work, we present a deep-learning-based super-resolution technique of confocal microscopy. We devise a two-channel attention network (TCAN), which takes advantage of both spatial representations and frequency contents to learn a more precise mapping from low-resolution images to high-resolution ones. This scheme is robust against changes in the pixel size and the imaging setup, enabling the optimal model to generalize to different fluorescence microscopy modalities unseen in the training set. Our algorithm is validated on diverse biological structures and dual-color confocal images of actin-microtubules, improving the resolution from ~ 230 nm to ~ 110 nm. Last but not least, we demonstrate live-cell super-resolution imaging by revealing the detailed structures and dynamic instability of microtubules.

Three-dimensional (3D) cultured tumor spheroid models, as one type of

Computed tomography (CT) allows for high lateral and axial resolution imaging of the endogenous structure of matter thanks to its large spatial frequency support and has been realized in X-ray and linear optical domain known as optical diffraction tomography (ODT). Here, we present the theoretical basis and experimental considerations for ODT of second-order nonlinear structures in weak scattering media. We have derived the relation between second harmonic wave and the anisotropic nonlinear tensor in spatial frequency domain under first-order Born approximation. Our results show that, under a plane wave illumination, the two dimensional (2D) spatial spectra of generated second harmonic complex field relates to the inverse lattice of nonlinear structure on Ewald sphere shells. The centers of the Ewald spheres are determined by 2 times wavevector of the incident fundamental wave and the radii are determined by the modulus of the second harmonic wavevector. More importantly, it shows that the 2D spatial spectra is a superposition of the Ewald spheres of different components of the anisotropic nonlinear tensor. We propose to solve the inverse problem by controlling the polarizations of the fundamental and second harmonic signal. We tested the feasibility of the proposed method using a numerical phantom and make some discussions on practical implementations, including angular scanning schemes, polarization detection and illumination profile for optimizing reconstruction region. Possessing high resolution, wide-field imaging and polarization-sensitive property, we believe that the proposed scheme would have important applications in nonlinear microscopy.

The algorithm used for reconstruction or resolution enhancement is one of the factors affecting the quality of super-resolution images obtained by fluorescence microscopy. Deep-learning-based algorithms have achieved state-of-the-art performance in super-resolution fluorescence microscopy and are becoming increasingly attractive. We firstly introduce commonly-used deep learning models, and then review the latest applications in terms of the network architectures, the training data and the loss functions. Additionally, we discuss the challenges and limits when using deep learning to analyze the fluorescence microscopic data, and suggest ways to improve the reliability and robustness of deep learning applications.

Stimulated emission depletion (STED) nanoscopy enables the visualization of subcellular organelles in unprecedented detail. However, reducing the power dependency remains one of the greatest challenges for STED imaging in living cells. Here, we propose a new method, called modulated STED, to reduce the demand for depletion power in STED imaging by modulating the information from the temporal and spatial domains. In this approach, an excitation pulse is followed by a depletion pulse with a longer delay; therefore, the fluorescence decay curve contains both confocal and STED photons in a laser pulse period. With time-resolved detection, we can remove residual diffraction-limited signals pixel by pixel from STED photons by taking the weighted difference of the depleted photons. Finally, fluorescence emission in the periphery of an excitation spot is further inhibited through spatial modulation of fluorescent signals, which replaced the increase of the depletion power in conventional STED. We demonstrate that the modulated STED method can achieve a resolution of < 100 nm in both fixed and living cells with a depletion power that is dozens of times lower than that of conventional STED, therefore, it is very suitable for long-term super-resolution imaging of living cells. Furthermore, the idea of the method could open up a new avenue to the implementation of other experiments, such as light-sheet imaging, multicolor and three-demensional (3D) super-resolution imaging.

Recently, CH3NH3PbX3 (X = Cl, Br, I) perovskites have been widely reported in photoelectric fields. However, the lower absorption efficiency and poor stability have restricted their further applications. In this work, we propose an absorber composed of CH3NH3PbI3 perovskite, silver nanowires (AgNWs) and TiO2 NWs. Using AgNWs and TiO2 NWs, the combined perovskite material had a maximum tenfold absorption enhancement than pure CH3NH3PbI3 perovskite, and the enhancement effect was higher than the recent report we carried out using silver nanowires (AgNWs), in which we obtained a maximum fivefold enhancement. In addition, the proposed complex showed good long-term stability and effectively prevented CH3NH3PbI3 perovskite from interacting with the active molecules in the air. Our work demonstrates that a synergistic effect was effectively utilized by AgNWs and TiO2 NWs and provides a facile method for absorption and stability improvement of CH3NH3PbI3 perovskites, thus broadening their photoelectric applications.

The rapid development of advanced optical imaging methods including stimulated emission depletion (STED) and fluorescence lifetime imaging microscopy (FLIM) has provided powerful tools for real-time observation of submicrometer biotargets to achieve unprecedented spatial and temporal resolutions. However, the practical imaging qualities are often limited by the performance of fluorescent probes, leading to unsatisfactory results. In particular, long-term imaging of nucleic acids in living cells with STED and FLIM remained desirable yet challenging due to the lack of competent probes combining targeting specificity, biocompatibility, low power requirement, and photostability. In this work, we rationally designed and synthesized a nanosized carbonized polymer dot (CPD) material, CPDs-3, with highly efficient and photostable emission for the super-resolution and fluorescence lifetime imaging of nucleic acids in living cells. The as-fabricated nanoprobe showed responsive emission properties upon binding with nucleic acids, providing an excellent signal-to-noise ratio in both spatial and temporal dimensions. Moreover, the characteristic saturation intensity value of CPDs-3 was as low as 0.68 mW (0.23 MW/cm2), allowing the direct observation of chromatin structures with subdiffraction resolution (90 nm) at very low excitation (

Low-level light therapy (LLLT) is emerging as a promising therapeutic approach to modulate the biochemical and molecular processes within living cells. LLLT is known to produce local and systemic effects; therefore, immune cells in local tissues or in the circulation are affected by light. However, this specific effect remains weakly explored. In this study, the effect of red (650 nm) and NIR (808 nm) light on phagocytosis (respiratory burst), cytokine expression, mitochondrial activity, ROS generation, Ca2+ influx and membrane depolarization in macrophages in vitro is investigated. Both the phagocytic capacity and adhesion of macrophages strongly (˜2.5 times) increased in the first hours after exposure to light in a dose-dependent manner. The light-evoked upregulation of phagocytosis is found to be less efficient than the maximal pharmacologically induced enhancement of ˜3.2 times. Also, red/NIR light reduces the production of pro-inflammatory cytokines and activates the secretion of anti-inflammatory cytokines by several times in activated macrophages. At the same time, the viability shows a biphasic dose response: it increases after irradiation with lower doses (0.3-1 J cm-2 ) and decreases after treatment with higher doses (18-30 J cm-2 ), which is apparently associated with the upregulation of ROS generation, followed by an increase in the mitochondrial activity.

Rationale: Acute kidney injury (AKI) is associated with aberrant generation of oxidative species and inflammation, leading to high mortality of in-hospitalized patients. Although N-acetylcysteine (NAC) showed positive effects in alleviating contrast-induced AKI, the clinical applications are strongly restrained due to the low bioavailability, low renal accumulation, short renal retention time, and high dosage-induced toxicity. Methods: We addressed the clinical dilemma of NAC by developing ultrasmall gold nanoclusters (1-2 nm) capped with NAC (denoted as Au NCs-NAC) as a nanozyme-based antioxidant defense system for AKI alleviation. Rhabdomyolysis-induced AKI mice model was developed, and the same dose of free NAC (as a control) and NAC onto Au NCs (Au NCs-NAC) was used for in vivo investigation of AKI restoration. Results: The as-developed gold nanozyme exhibited high bioavailability and good physicochemical stability as compared to NAC. Meanwhile, Au NCs-NAC showed broad-spectrum antioxidant activity of Au NCs-NAC, offering in vitro renoprotective effects, as well as macrophages by relieving inflammation under hydrogen peroxide or lipopolysaccharide stimulation. Notably, owing to the smaller size than kidney threshold (5.5 nm), Au NCs-NAC displayed preferential renal enrichment (< 2 h) and longer retention (> 24 h) in AKI mice as revealed by fluorescence imaging, thereby largely enhancing the restoration of renal function in AKI mice than free NAC by protecting the kidneys from oxidative injury and inflammation without systemic toxicity, as demonstrated by tissues staining, inflammatory cytokines and biomarkers detection, and mice survival rate. Conclusion: Owing to the synergistic anti-inflammatory/antioxidative effects, and enhanced bioavailability and renal accumulation/retention, Au NCs-NAC displayed far superior therapeutic performance than NAC alone. This work will facilitate the development of high-performance antioxidative nanoplatforms, as well as overcome the clinical limitations of small molecular drugs for AKI treatment and other inflammatory diseases.

The conventional approach in cancer nanomedicine involves advanced drug nanocarriers delivering preloaded therapeutics to targeted tumor sites to maximize drug efficiency. However, both cancer drugs and nanocarriers inevitably produce side effects and systemic toxicity. Herein, hemoglobin nanocrystals (HbC) as drug-free theranostic nanoformulations with the tumor microenvironment (TME) activated diagnostic and therapeutic abilities towards colon tumors are introduced. HbC can release Fe