An Extended NIR-II Superior Imaging Window from 1500 to 1900 nm for High-Resolution In Vivo Multiplexed Imaging Based on Lanthanide Nanocrystals

High-resolution in vivo optical multiplexing in second near-infrared window (NIR-II, 1000–1700 nm) is vital to biomedical research. Presently, limited by bio-tissue scattering, only luminescent probes located at NIR-IIb (1500–1700 nm) window can provide high-resolution in vivo multiplexed imaging. However, the number of available luminescent probes in this narrow NIR-IIb region is limited, which hampers the available multiplexed channels of in vivo imaging. To overcome the above challenges, through theoretical simulation we expanded the conventional NIR-IIb window to NIR-II long-wavelength (NIR-II-L, 1500–1900 nm) window on the basis of photon-scattering and water-absorption. We developed a series of novel lanthanide luminescent nanoprobes with emission wavelengths from 1852 nm to 2842 nm. NIR-II-L nanoprobes enabled high-resolution in vivo dynamic multiplexed imaging on blood vessels and intestines, and provided multi-channels imaging on lymph tubes, tumors and intestines. The proposed NIR-II-L probes without mutual interference are powerful tools for high-contrast in vivo multiplexed detection, which holds promise for revealing physiological process in living body.     

稀土纳米材料在高分辨活体成像及诊疗中的应用

荧光成像具有时空分辨率高、 反馈快、 非侵入和无电离辐射等优点, 是一种重要的生物成像技术. 与传统用于荧光成像的可见光和近红外一区(NIR-I, 600~950 nm)相比, 近红外二区(NIR-Ⅱ, 1000~1700 nm)窗口具有低生物组织散射系数和低生物自发荧光, 采用NIR-Ⅱ光进行活体荧光成像能有效提高成像的分辨率、 信噪比和穿透深度. 稀土纳米颗粒(RENPs)具有大斯托克斯位移、 高化学稳定性、 可调的荧光寿命以及较窄的发射带, 是一种重要的荧光成像探针. 近年来, 一系列具有优异的NIR-Ⅱ发光性能的稀土纳米材料被用于高分辨活体荧光成像. 本文综合评述了近年来RENPs用于高分辨活体成像及诊疗中的研究进展, 概述了RENPs的掺杂调控、 基质晶格选择和复合敏化等NIR-Ⅱ发光增强策略, 介绍了其在多种生物医学场景中的靶向聚集、 荧光传感和疾病治疗等功能, 并总结了其在多路成像、 多模态成像和疾病诊疗中的应用. 最后, 简要分析了RENPs在未来生物医学应用中面临的挑战和发展的方向.     

Photonics-on-a-chip: recent advances in integrated waveguides as enabling detection elements for real-world, lab-on-a-chip biosensing applications

By leveraging advances in semiconductor microfabrication technologies, chip-integrated optical biosensors are poised to make an impact as scalable and multiplexable bioanalytical measurement tools for lab-on-a-chip applications. In particular, waveguide-based optical sensing technology appears to be exceptionally amenable to chip integration and miniaturization, and, as a result, the recent literature is replete with examples of chip-integrated waveguide sensing platforms developed to address a wide range of contemporary analytical challenges. As an overview of the most recent advances within this dynamic field, this review highlights work from the last 2-3 years in the areas of grating-coupled, interferometric, photonic crystal, and microresonator waveguide sensors. With a focus towards device integration, particular emphasis is placed on demonstrations of biosensing using these technologies within microfluidically controlled environments. In addition, examples of multiplexed detection and sensing within complex matrices--important features for real-world applicability--are given special attention.     

Spatially resolved analyte mapping with time-of-flight optical sensors

Simultaneous real-time acquisition of analytical data from several locations is attractive in a variety of applications. This brief review traces the evolution of approaches to such measurements. Greatest emphasis is placed on optical time-of-flight chemical detection when signals are multiplexed from several point sensors or when the measurements are taken along the length of a single continuous extended-length ‘distributed' sensing element. The use of sensors featuring extended-length continuous chemically sensitive optical fibers offers detection arrangements for which there is no counterpart in conventional chemical sensor technologies.     

Surface enhanced Raman scattering for multiplexed detection

The multiplexed detection of biological analytes from complex mixtures is of crucial importance for the future of intelligent management and detection of disease. This review focuses on recent advances in the use of surface enhanced Raman scattering (SERS) spectroscopy as an analytical technique that can deliver multiplexed detection for a variety of biological target in increasingly complex media. The use of SERS has developed from the multipelxed detection of custom dye molecules to biomolecules such as DNA and proteins. Recent work has also shown the capability of SERS multiplexing for in vivo as well as in vitro applications.     

Molecular luminescence imaging

Imaging techniques relying on different luminescence processes are powerful bioanalytical tools for life sciences. They are used in multiplexed quantitative assays in different analytical formats and to assess the spatial distribution of a given target molecule, chemical or biochemical process in macro- and microsamples, including the in vivo evaluation of biological and pathological processes. Here, recent progresses in luminescence imaging techniques are described, including new labels for fluorescence, time-resolved fluorescence and bio-chemiluminescence, multiplexed imaging microscopy techniques and whole-body luminescence imaging in live animals.     

N-glycosylation analysis of mouse immunoglobulin G isolated from dried blood spots

The association of immunoglobulin G (IgG) glycosylation changes with various human diseases and physiological conditions is well established. Since the mechanistical explanation of the regulation of IgG glycosylation and its functional role in these various states is still missing, the eyes of the biomedical community are now turned towards animal models, which enable intervention studies necessary for conclusions on causality. Mice are recognized and used as a good experimental model for human IgG glycosylation. However, smaller blood volumes, low IgG concentrations at young ages (which are most often used in mice experiments) and multiple sampling protocols during the course of longitudinal studies would profit from a robust workflow for mouse IgG glycome analysis from minute amounts of starting material, collected through a simple sampling procedure. For this purpose, we have developed a protocol for analysis of total N-glycans of IgG isolated from mouse dried blood spots (DBS), which we report here. We show that mouse DBS are a good source of material for IgG N-glycan analysis by multiplexed capillary gel electrophoresis with laser-induced fluorescence (xCGE-LIF).     

Automated analytical microarrays: a critical review

Microarrays provide a powerful analytical tool for the simultaneous detection of multiple analytes in a single experiment. The specific affinity reaction of nucleic acids (hybridization) and antibodies towards antigens is the most common bioanalytical method for generating multiplexed quantitative results. Nucleic acid-based analysis is restricted to the detection of cells and viruses. Antibodies are more universal biomolecular receptors that selectively bind small molecules such as pesticides, small toxins, and pharmaceuticals and to biopolymers (e.g. toxins, allergens) and complex biological structures like bacterial cells and viruses. By producing an appropriate antibody, the corresponding antigenic analyte can be detected on a multiplexed immunoanalytical microarray. Food and water analysis along with clinical diagnostics constitute potential application fields for multiplexed analysis. Diverse fluorescence, chemiluminescence, electrochemical, and label-free microarray readout systems have been developed in the last decade. Some of them are constructed as flow-through microarrays by combination with a fluidic system. Microarrays have the potential to become widely accepted as a system for analytical applications, provided that robust and validated results on fully automated platforms are successfully generated. This review gives an overview of the current research on microarrays with the focus on automated systems and quantitative multiplexed applications. Figure MCR 3: A fully automated chemiluminescence microarray reader for analytical microarrays     

Potential of label-free detection in high-content-screening applications

The classical approach of high-content screening (HCS) is based on multiplexed, functional cell-based screening and combines several analytical technologies that have been used before separately to achieve a better level of automation (scale-up) and higher throughput. New HCS methods will help to overcome the bottlenecks, e.g. in the present development chain for lead structures for the pharmaceutical industry or during the identification and validation process of new biomarkers. In addition, there is a strong need in analytical and bioanalytical chemistry for functional high-content assays which can be provided by different hyphenated techniques. This review discusses the potential of a label-free optical biosensor based on reflectometric interference spectroscopy (RIfS) as a bridging technology for different HCS approaches. Technical requirements of RIfS are critically assessed by means of selected applications and compared to the performance characteristics of surface plasmon resonance (SPR) which is currently the leading technology in the area of label-free optical biosensors.     

无机半导体纳米晶光学性质的设计、调控及其生物医学应用

半导体纳米晶由于其丰富的能带结构和光学性质,在光电器件和生物医学应用等领域展现出了广阔的应用前景,且在过去的几十年中得到了广泛关注.因此,对其光学性质进行理性设计和精确调控具有重要的研究意义.本文简要综述了本研究组近年来在不同能带隙的无机半导体纳米晶的可控制备技术以及利用DNA纳米技术和蛋白质自组装手段构建具有特异光学性质的纳米结构等方面的相关研究工作,最后对这些纳米晶和纳米结构的独特光学性质及其在生物医学领域的应用研究进行了总结.     

Diagnostic prospects and preclinical development of optical technologies using gold nanostructure contrast agents to boost endogenous tissue contrast

Numerous developments in optical biomedical imaging research utilizing gold nanostructures as contrast agents have advanced beyond basic research towards demonstrating potential as diagnostic tools; some of which are translating into clinical applications. Recent advances in optics, lasers and detection instrumentation along with the extensive, yet developing, knowledge-base in tailoring the optical properties of gold nanostructures has significantly improved the prospect of near-infrared (NIR) optical detection technologies. Of particular interest are optical coherence tomography (OCT), photoacoustic imaging (PAI), multispectral optoacoustic tomography (MSOT), Raman spectroscopy (RS) and surface enhanced spatially offset Raman spectroscopy (SESORS), due to their respective advancements. Here we discuss recent technological developments, as well as provide a prediction of their potential to impact on clinical diagnostics. A brief summary of each techniques'' capability to distinguish abnormal (disease sites) from normal tissues, using endogenous signals alone is presented. We then elaborate on the use of exogenous gold nanostructures as contrast agents providing enhanced performance in the above-mentioned techniques. Finally, we consider the potential of these approaches to further catalyse advances in pre-clinical and clinical optical diagnostic technologies.

Optical biomedical imaging research utilising gold nanostructures as contrast agents has advanced beyond basic science, demonstrating potential in various optical diagnostic tools; some of which are currently translating into clinical applications.     

贵金属基纳米酶的研究进展

贵金属纳米材料在纳米尺度具有独特的光学、 电学性质及优异的催化性能, 是一类重要的功能纳米材料. 基于贵金属材料的纳米酶研究是贵金属纳米材料在生物医学领域的一个前沿研究方向. 贵金属基纳米酶具有特殊的光学性质、 较好的化学稳定性、 可调控的类酶活性及良好的生物相容性, 是目前纳米生物医学领域的热点研究材料. 本文总结了贵金属基纳米酶的活性种类、 活性机理、 活性调控以及在生物医学等领域的潜在应用.     

Recent developments in detection for microfluidic systems

Microfluidic systems have become more and more important in the field of analytical chemistry. Detection methods on these microsystems are essential for the identification and quantification of chemical species that are being analyzed. This review concentrates on the latest developments of optical detection methods and mass spectrometry in conjunction with microfluidic systems. Electrochemical methods are discussed in another review in the same issue of this journal. Within the optical detection section, topics such as multiplexed detection and the use of waveguides are discussed. Within the discussion of mass spectrometry, the main focus is on electrospray emitters as interfaces between microsystem and spectrometer. Apart from optical detection and mass spectrometry, other techniques such as flame ionization and nuclear magnetic resonance are also mentioned.     

Electrochemical Signal Substance for Multiplexed Immunosensing Interface Construction: A Mini Review

Appropriate labeling method of signal substance is necessary for the construction of multiplexed electrochemical immunosensing interface to enhance the specificity for the diagnosis of cancer. So far, various electrochemical substances, including organic molecules, metal ions, metal nanoparticles, Prussian blue, and other methods for an electrochemical signal generation have been successfully applied in multiplexed biosensor designing. However, few works have been reported on the summary of electrochemical signal substance applied in constructing multiplexed immunosensing interface. Herein, according to the classification of labeled electrochemical signal substance, this review has summarized the recent state-of-art development for the designing of electrochemical immunosensing interface for simultaneous detection of multiple tumor markers. After that, the conclusion and prospects for future applications of electrochemical signal substances in multiplexed immunosensors are also discussed. The current review can provide a comprehensive summary of signal substance selection for workers researched in electrochemical sensors, and further, make contributions for the designing of multiplexed electrochemical immunosensing interface with well signal.     

Layer double hydroxides (LDHs)- based electrochemical and optical sensing assessments for quantification and identification of heavy metals in water and environment samples: A review of status and prospects

One of the most severe environmental problems is heavy metal contamination, putting the world's sustainability at risk. Much effort has been put into developing sensors that can be taken anywhere to detect the environmental effects of heavy metals. Sensitivity, selectivity, multiplexed detection ability, and mobility enhance significantly when nanoparticles and nanostructures are incorporated into sensors. LDHs (layered double hydroxides) have gotten much attention in analytical chemistry in recent years because of their benefits, including their large specific surface area, ease of synthesis, low cost, and high catalytic efficiency and biocompatibility. LDHs are often manufactured as nanomaterial composites or created with specialized three-dimensional structures depending on the application. However, in these settings, LDHs (as color indicators, extracting sorbents, and electrochemical sensing) are usually restricted. Upcoming signs of progress and development possibilities of LDHs in analytical chemistry are reviewed in this paper to assist overcome these problems. Furthermore, the approaches used in the design of LDHs, including structural aspects, are defined and assessed in preparation for future analytical applications. The latest advances in optical and electrochemical sensors to detect heavy metals are described in this review. The sorts and characteristics of LDHs will be explored first. We will then go into microelectrode (or nanoelectrode) arrays, nanoparticle-modified electrodes, and microfluidic optical and electrochemical sensing assays in detail. This paper also discusses design strategies for LDH-based nanostructured sensors and the advantages of using nanomaterials and nanostructures.     

Development of a multiplexed microbioreactor system for high-throughput bioprocessing

A multiplexed microbioreactor system for parallel operation of multiple microbial fermentation is described. The system includes miniature motors for magnetic stirring of the microbioreactors and optics to monitor the fermentation parameters optical density (OD), dissolved oxygen (DO), and pH, in-situ and in real time. The microbioreactors are fabricated out of poly(methylmethacrylate)(PMMA) and poly(dimethylsiloxane)(PDMS), and have a working volume of 150 microl. Oxygenation of the cells occurs through a thin PDMS membrane at the top of the reactor chamber. Stirring is achieved with a magnetic spin bar in the reactor chamber. Parallel microbial fermentations with Escherichia coli are carried out in four stirred microbioreactors and demonstrate the reproducible performance of the multiplexed system. The profiles for OD, DO, and pH compare favourably to fermentations performed in bioreactor systems with multiple bench-scale reactors. Finally, the multiplexed system is used to compare two different reactor designs, demonstrating that the reproducibility of the system permits the quantification of microbioreactor performance.     

核酸适配体及其在化学领域的相关应用

核酸适配体是一小段经体外筛选得到的寡核苷酸序列,能与相应的配体进行高亲和力和强特异性的结合,它的出现为化学生物学界和生物医学界提供了一种新的高效快速识别的研究平台,并在许多方面展示了良好的应用前景。本文从核算适配体的性质和体外筛选过程等方面出发,着重综述了核算适配体的化学修饰方法及其在分析化学和酶化学中应用的研究进展。     

Color-encoded microcarriers based on nano-silicon dioxide film for multiplexed immunoassays

Multiplexed analysis allows researchers to obtain high-density information with minimal assay time, sample volume and cost. Currently, microcarrier or particle-based approaches for multiplexed analysis involve complicated or expensive encoding and decoding processes. In this paper, a novel optical encoding technique based on nano-silicon dioxide film is presented. Microcarriers composed of thermally grown silicon dioxide (SiO(2)) film and monocrystalline silicon (Si) substrate were fabricated. The nano-silicon dioxide film exhibited unique surface color by low-coherence interference. Hence the colors can be used for encoding at least 100 microcarriers loaded with films of different thickness. We demonstrated that color-encoded microcarriers loaded with antigens could be used for multiplexed immunoassays to detect goat anti-human IgG, goat anti-mouse IgG and goat anti-rabbit IgG, with fluorescent detection as the interrogating approach. This microcarrier-based method also exhibited improved analytical performance compared with a microarray technique. This approach will provide new opportunities for multiplexed target assay development.     

Universal Super‐Resolution Multiplexing by DNA Exchange

Super‐resolution microscopy allows optical imaging below the classical diffraction limit of light with currently up to 20× higher spatial resolution. However, the detection of multiple targets (multiplexing) is still hard to implement and time‐consuming to conduct. Here, we report a straightforward sequential multiplexing approach based on the fast exchange of DNA probes which enables efficient and rapid multiplexed target detection with common super‐resolution techniques such as (d)STORM, STED, and SIM. We assay our approach using DNA origami nanostructures to quantitatively assess labeling, imaging, and washing efficiency. We furthermore demonstrate the applicability of our approach by imaging multiple protein targets in fixed cells.     

基于低相干干涉的透镜厚度测量及生物影像研究进展

低相干光干涉测量技术作为重要的非接触测量方法之一,由于具有结构简单、测量速度快及分辨率高的优点,在光学系统的非接触测量及生物医学影像等前沿领域具有广阔的应用前景。本文对透镜组的光学元件中心厚度及空气间隔的非接触测量技术进行了总结,简述了基于OCT的生物医学影像方面研究现状,重点论述基于低相干光干涉法测量原理和研究进展,对比分析相关研究方法的优缺点和创新之处,并从系统结构和应用范围的角度对低相干干涉测量技术的发展趋势作了展望。