in vivo and in situ robust quantitative optical biopsy of hepatocellular carcinoma and metastasis based on two-photon autofluorescence imaging.

Two-photon autofluorescence (TPAF) imaging is able to offer precise cellular metabolic information with high spatiotemporal resolution, making it a promising biopsy tool. The technique is greatly hampered by the complexity of either the optical system or data processing. Here, the excitation wavelength was optimized to simultaneously excite both flavin adenine dinucleotide and nicotinamide adenine dinucleotide and eliminate the unexpected TPAF. The optical redox ratio (ORR) images were robustly achieved without additional calibration under the optimized single-wavelength excitation. The in vitro, ex vivo, and in vivo biopsy by the TPAF method were systematically studied and compared using hepato-cellular carcinoma and metastasis as examples. It was demonstrated that the proposed TPAF method simplified the optical system, improved the robustness of ORR, and enabled early-stage cancer diagnosis, showing distinguished advantages as compared with previous methods.

Biocompatible polymer optical fiber with a strongly scattering spherical end for interstitial photodynamic therapy.

Interstitial photodynamic therapy (I-PDT), which utilizes optical fibers to deliver light for photosensitizer excitation and the elimination of penetration depth limitation, is a promising modality in the treatment of deeply seated tumors or thick tumors. Currently, the excitation domain of the optical fiber is extremely limited, restricting PDT performance. Here, we designed and fabricated a biocompatible polymer optical fiber (POF) with a strongly scattering spherical end (SSSE) for I-PDT applications, achieving an increased excitation domain and consequently excellent in vitro and in vivo therapeutical outcomes. The POF, which was drawn using a simple thermal drawing method, was made of polylactic acid, ensuring its superior biocompatibility. The excitation domains of POFs with different ends, including flat, spherical, conical, and strongly scattering spherical ends, were analyzed and compared. The SSSE was achieved by introducing nanopores into a spherical end, and was further optimized to achieve a large excitation domain with an even intensity distribution. The optimized POF enabled outstanding therapeutic performance of I-PDT in in vitro cancer cell ablation and in vivo anticancer therapy. All of its notable optical features, including low transmission/bending loss, superior biocompatibility, and a large excitation domain with an even intensity distribution, endow the POF with great potential for clinical I-PDT applications.

In Vivo Flow Cytometry.

In vivo flow cytometry (IVFC) was first designed to detect circulating cells in a mouse ear. It allows real-time monitoring of cells in peripheral blood with no need to draw blood. The IVFC field has made great progress during the last decade with the development of fluorescence, photoacoustic, and multiphoton microscopy. Moreover, the application of IVFC is no longer restricted to circulating cells. IVFC based on fluorescence and photoacoustic are most widely applied in biomedical research. Methods based on fluorescence are often used for object monitoring in superficial vessels, while methods based on photoacoustics have an advantage of label-free monitoring in deep vessels. In this chapter, we introduce technical points and key applications of IVFC. We focus on the principles, labeling strategies, sensitivity, and biomedical applications of the technology. In addition, we summarize this chapter and discuss important research directions of IVFC in the future.

Fundamentals of Optical Imaging.

Optical imaging, which possesses noninvasive and high-resolution features for biomedical imaging, has been used to study various biological samples, from in vitro cells, ex vivo tissue, to in vivo imaging of living organism. Furthermore, optical imaging also covers a very wide scope of spatial scale, from submicron sized organelles to macro-scale live biological samples, enabling it a powerful tool for biomedical studies. Before introducing these superior optical imaging methods to researchers, first of all, it is necessary to present the basic concept of light-matter interactions such as absorption, scattering, and fluorescence, which can be used as the imaging contrast and also affect the imaging quality. And then the working mechanism of various imaging modalities including fluorescence microscopy, confocal microscopy, multiphoton microscopy, super-resolution microscopy, optical coherence tomography (OCT), diffuse optical tomography (DOT), etc. will be presented. Meanwhile, the main features and typical bioimaging applications of these optical imaging technologies are discussed. Finally, the perspective of future optical imaging methods is presented. The aim of this chapter is to introduce the background and principle of optical imaging for grasping the mechanism of advanced optical imaging modalities introduced in the following chapters.

Flexible porphyrin doped polymer optical fibers for rapid and remote detection of trace DNT vapor.

With the rapid growth of anti-terrorist activities worldwide, it becomes an emerging requirement to rapidly and accurately detect hidden explosive threats. However, the safety issue during the explosive material detection, e.g. unexpected explosion, is still an insurmountable challenge. In this study, we design and mass-produce a novel kind of flexible 5,10,15,20-tetrakis(4-aminophenyl)porphyrin doped polymer optical fiber (PPOF) for rapid and accurate detection of trace 2,4-dinitrotoluene (DNT) vapor based on the DNT induced florescence quenching mechanism. The influence of doping concentration, bending, and temperature on the sensing performance is investigated. PPOF shows immunity to bending, enabling it to work in a harsh environment. It is experimentally demonstrated that the limit of detection and response time of the proposed PPOF could reach around 120 ppb and 3 minutes, respectively, which make it much better than other techniques. Owning to its inherent advantages including low-cost, remote-control capability, and compatibility with optical communication networking, PPOF can be constructed the quasi-distributed sensing networking of explosive matters in the future, providing a new strategy for anti-terrorism.

Recent advances in copper sulphide-based nanoheterostructures.

Due to the high mobility of copper ions in numerous structurally-related phases, copper sulphide (Cu2-xS, 0 ≤x≤ 1) has been widely used as a starting template to fabricate various heterostructures via cation exchange. Such nanoheterostructures can possess unique combinations of physical properties that could be useful in diverse applications. Controllable methods of fabricating copper sulphide nanoheterostructures of increasing complexity have been rapidly emerging over the past few years. In this tutorial review, we discuss recent progress in heterostructure fabrication methods using copper sulphide. We primarily focus on important reports of cation exchange-based approaches and then summarize some key emerging applications that can employ these copper-sulphide-based nanoheterostructures.

Tunable hybridization induced transparency for efficient terahertz sensing.

Hybridization induced transparency (HIT) resulting from the coupling between the material absorption resonance and the artificial structure (metamaterial) resonance provides an effective means of enhancing the sensitivity in the terahertz spectroscopic technique-based sensing applications. However, the application of this method is limited by the versatility to the samples with different volumes, because the samples usually have a refractive index larger than unity and their presence with different thicknesses will lead to a shift of the structure resonance, mismatching the material absorption. In this work, we demonstrate that by using InSb coupled rod structures, whose electromagnetic response in the terahertz band can be easily controlled by using ambient parameters like the temperature or magnetic field, the HIT effect can be easily tuned so that without the needs to change the rod geometry, one can realize efficient terahertz sensing with different sample thickness.

Controlled excitation of higher radial order whispering gallery modes with metallic diffraction grating.

Metallic diffraction grating coupler is investigated for controlled excitation of whispering gallery modes (WGMs) of different radial orders. Based on effective mode index calculations and finite difference time domain method, it is found that higher radial order WGMs can be separated from the fundamental modes by sending them into the opposite propagation direction. By phase-matching designs, the metallic diffraction grating provides extra freedom to switch propagation directions, and is able to selectively enhance or suppress different radial-order WGMs. Such structure offers a simple and practical configuration for various WGM applications including liquid sensing, band pass filtering and fiber lasers.

Power transfer mechanism of metallic grating coupled whispering gallery microsphere resonator.

The power transfer mechanism of the Fano resonance in a metallic grating coupled whispering gallery microsphere resonator is both theoretically investigated and experimentally verified. Based on temporal coupled mode theory, a two-stage cascaded scattering model is formulated to account for the power interaction of multiple resonances in the system. The modeled results show agreement with experimental measurements and are able to explain several counter-intuitive observations in the spectra. The coupling efficiency of the grating coupler is evaluated at critical coupling by approximating symmetric resonance line-shape with grating design. Based on the modeled results, it is found that the coupling efficiency can be further improved by enhancing diffraction efficiency of the first-order diffractions.

Reflective liquid level sensor based on modes conversion in thin-core fiber incorporating titled fiber Bragg grating.

A simple and compact reflective liquid level sensor based on modes conversion in the thin-core fiber incorporating one tilted fiber Bragg grating (TFBG) is proposed and experimentally demonstrated. A piece of thin-core fiber containing one TFBG ensures the modes conversion between the core mode and cladding modes. The external liquid can induce the cladding modes covert to the radiation modes and lead to the decrement of the collected cladding modes power, then the liquid level can be measured from the collected cladding modes power. The modes conversion in the proposed structure is theoretically analyzed. The experimental results show the high liquid level sensitivity and temperature immunity of the proposed sensor, and its significant advantage is that the measurement range is not limited to the length of the TFBG itself.

Simple and compact reflective refractometer based on tilted fiber Bragg grating inscribed in thin-core fiber.

A simple and compact reflective refractometer based on a tilted fiber Bragg grating (TFBG) inscribed in an ultra-high photon-sensitive thin-core fiber is proposed and experimentally demonstrated. The reflective refractometer utilizes a short piece of thin-core fiber containing one TFBG to ensure the recoupling of cladding modes. The reflection spectra occur in two well-defined wavelength bands that correspond to the Bragg core mode and cladding modes, respectively. It is found that the power of the collected cladding modes changes with the external refractive index (RI), while that of the Bragg core mode remains unaffected and can be used as the temperature reference. High RI sensitivity and temperature immunity of the proposed reflective refractometer are experimentally achieved.

Boosted photo-immunotherapy via near-infrared light excited phototherapy in tumor sites and photo-activation in sentinel lymph nodes.

Phototherapy is a promising modality that could eradicate tumor and trigger immune responses via immunogenic cell death (ICD) to enhance anti-tumor immunity. However, due to the lack of deep-tissue-excitable phototherapeutic agents and appropriate excitation strategies, the utility of phototherapy for efficient activation of the immune system is challenging. Herein, we report functionalized ICG nanoparticles (NPs) with the capture capability of tumor-associated antigens (TAAs). Under near-infrared (NIR) light excitation, the ICG NPs exhibited high-performance phototherapy, i.e., synergistic photothermal therapy and photodynamic therapy, thereby efficiently eradicating primary solid tumor and inducing ICD and subsequently releasing TAAs. The ICG NPs also captured TAAs and delivered them to sentinel lymph nodes, and then the sentinel lymph nodes were activated with NIR light to trigger efficient T-cell immune responses through activation of dendritic cells with the assistance of ICG NP generated reactive oxygen species, inhibiting residual primary tumor recurrence and controlling distant tumor growth. The strategy of NIR light excited phototherapy in tumor sites and photo-activation in sentinel lymph nodes provides a powerful platform for active immune systems for anti-tumor photo-immunotherapy.

Data-driven coordinated attention deep learning for high-fidelity brain imaging denoising and inpainting.

Deep learning offers promise in enhancing low-quality images by addressing weak fluorescence signals, especially in deep in vivo mouse brain imaging. However, current methods struggle with photon scarcity and noise within in vivo deep mouse brains, and often neglecting tissue preservation. In this study, we propose an innovative in vivo cortical fluorescence image restoration approach, combining signal enhancement, denoising, and inpainting. We curated a deep brain cortical image dataset and developed a novel deep brain coordinate attention restoration network (DeepCAR), integrating coordinate attention with optimized residual networks. Our method swiftly and accurately restores deep cortex images exceeding 800 µm, preserving small-scale tissue structures. It boosts the peak signal-to-noise ratio (PSNR) by 6.94 dB for weak signals and 11.22 dB for large noisy images. Crucially, we validate the effectiveness on external datasets with diverse noise distributions, structural features compared to those in our training data, showcasing real-time high-performance image restoration capabilities.

General Post-Regulation Strategy of AIEgens' Photophysical Properties for Intravital Two-Photon Fluorescence Imaging.

Fluorogens with aggregation-induced emission (AIEgens) are promising agents for two-photon fluorescence (TPF) imaging. However, AIEgens' photophysical properties are fixed and unoptimizable once synthesized. Therefore, it is urgent and meaningful to explore an efficient post-regulation strategy to optimize AIEgens' photophysical properties. Herein, a general and efficient post-regulation strategy is reported. By simply tuning the ratio of inert AIEgens within binary nanoparticles (BNPs), the fluorescence quantum yield and two-photon absorption cross-section of functional AIEgens are enhanced by 8.7 and 5.4 times respectively, which are not achievable by conventional strategies, and the notorious phototoxicity is almost eliminated. The experimental results, theoretical simulation, and mechanism analysis demonstrated its feasibility and generality. The BNPs enabled deep cerebrovascular network imaging with ≈1.10 mm depth and metastatic cancer cell detection with single-cell resolution. Furthermore, the TPF imaging quality is improved by the self-supervised denoising algorithm. The proposed binary molecular post-regulation strategy opened a new avenue to efficiently boost the AIEgens' photophysical properties and consequently TPF imaging quality.

Superior biocompatible carbon dots for dynamic fluorescence imaging of nucleoli in living cells.

The nucleolus is a newly developed and promising target for cancer diagnosis and therapy, and its imaging is extremely significant for fundamental research and clinical applications. The unique feature, i.e., high resolution at the subcellular level, makes the fluorescence imaging method a powerful tool for nucleolus imaging. However, the fluorescence imaging of nucleoli in living cells is restricted by the limited availability of fluorescent agents with specific nucleolus-targeting capability and superior biocompatibility. Here, promising carbon dots (CDs) with intrinsic nucleolus-targeting capability were synthesized, characterized and employed for dynamic fluorescence imaging of nucleoli in living cells. The CDs exhibit a high fluorescence quantum yield of 0.2, excellent specificity and photostability, and superior biocompatibility, which were systematically demonstrated at the gene, cellular and animal levels and confirmed by their biological effects on embryonic development. All these features enabled CDs to light up the nucleoli for a long time with a high signal-to-noise ratio in living cells and monitor the nucleolar dynamics of malignant cells in camptothecin (CPT) based chemotherapy. Their excellent optical and biological features as well as general nucleolus-targeting capability endow CDs with great potential for future translational research.

Advanced flow cytometry for biomedical applications.

Flow cytometry (FC) is a versatile tool with excellent capabilities to detect and measure multiple characteristics of a population of cells or particles. Notable advancements in in vivo photoacoustic FC, coherent Raman FC, microfluidic FC, and so on, have been achieved in the last two decades, which endows FC with new functions and expands its applications in basic research and clinical practice. Advanced FC broadens the tools available to researchers to conduct research involving cancer detection, microbiology (COVID-19, HIV, bacteria, etc.), and nucleic acid analysis. This review presents an overall picture of advanced flow cytometers and provides not only a clear understanding of their mechanisms but also new insights into their practical applications. We identify the latest trends in this area and aim to raise awareness of advanced techniques of FC. We hope this review expands the applications of FC and accelerates its clinical translation.

Nonlinear fiber-optic strain sensor based on four-wave mixing in microstructured optical fiber.

We demonstrate a nonlinear fiber-optic strain sensor, which uses the shifts of four-wave mixing Stokes and anti-Stokes peaks caused by the strain-induced changes in the structure and refractive index of a microstructured optical fiber. The sensor thus uses the inherent nonlinearity of the fiber and does not require any advanced postprocessing of the fiber. Strain sensitivity of -0.23 pm/µÎµ is achieved experimentally and numerical simulations reveal that for the present fiber the sensitivity can be increased to -4.46 pm/µÎµ by optimizing the pump wavelength and power.

The stiffness-dependent tumor cell internalization of liquid metal nanoparticles.

The properties of nanoparticle (NP) carriers, such as size, shape and surface state, have been proven to dramatically affect their uptake by tumor cells, thereby influencing and determining the effect of nanomedicine on tumor theranostics. However, the effect of the stiffness of NPs on their cellular internalization remains unclear, especially for circumstances involving active or passive NP targeting. In this work, we constructed eutectic gallium indium liquid metal NPs with the same particle size, shape and surface charge properties but distinct stiffness via tailoring the surface oxidation and silica coating. It has been found that the softer NPs would be endocytosed much slower than their stiffer counterparts in the presence of specific ligand-receptor interaction. Interestingly, once the interaction is eliminated, softer NPs are internalized faster than the stiffer ones. Based on experimental observations and theoretical verification, we demonstrate that this phenomenon is mainly caused by varying degrees of deformation of soft NPs induced by ligand-receptor interactions. Such a finding of the stiffness effect of NPs implies great potential for fundamental biomedical applications, such as the rational design of nanomedicines.

Light amplified oxidative stress in tumor microenvironment by carbonized hemin nanoparticles for boosting photodynamic anticancer therapy.

Photodynamic therapy (PDT), which utilizes light excite photosensitizers (PSs) to generate reactive oxygen species (ROS) and consequently ablate cancer cells or diseased tissue, has attracted a great deal of attention in the last decades due to its unique advantages. However, the advancement of PDT is restricted by the inherent characteristics of PS and tumor microenvironment (TME). It is urgent to explore high-performance PSs with TME regulation capability and subsequently improve the therapeutic outcomes. Herein, we reported a newly engineered PS of polymer encapsulated carbonized hemin nanoparticles (P-CHNPs) via a facile synthesis procedure for boosting photodynamic anticancer therapy. Solvothermal treatment of hemin enabled the synthesized P-CHNPs to enhance oxidative stress in TME, which could be further amplified under light irradiation. Excellent in vitro and in vivo PDT effects were achieved due to the improved ROS (hydroxyl radicals and singlet oxygen) generation efficiency, hypoxia relief, and glutathione depletion. Moreover, the superior in vitro and in vivo biocompatibility and boosted PDT effect make the P-CHNPs a potential therapeutic agent for future translational research.

Optical fiber relative humidity sensor based on FBG incorporated thin-core fiber modal interferometer.

A new fiber-optic relative humidity (RH) sensor based on a thin-core fiber modal interferometer (TCFMI) with a fiber Bragg grating (FBG) in between is presented. Poly (N-ethyl-4-vinylpyridinium chloride) (P4VP·HCl) and poly (vinylsulfonic acid, sodium salt) (PVS) are layer-by-layer deposited on the side surface of the sensor for RH sensing. The fabrication of the sensing nanocoating is characterized by using UV-vis absorption spectroscopy, quartz crystal microbalance (QCM) and scanning electron microscopy (SEM). The incorporation of FBG in the middle of TCFMI can compensate the cross sensitivity of the sensor to temperature. The proposed sensor can detect the RH with resolution of 0.78% in a large RH range at different temperatures. A linear, fast and reversible response has been experimentally demonstrated.