| 单颗粒光散射显微成像技术在生化医药分析中的应用 |
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| 基金项目:重庆市自然科学基金资助项目(cstc2019jcyj-msxmX0279)；中央高校业务费优青培育项目(XDJK2019AC002)；药学国家级实验教学中心(西南大学)大学生创新训练项目 |
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| 中文摘要:等离子体纳米颗粒(PNPs)具有体积小、易表面修饰、生物相容性好、毒性低等优点，在生物传感、生物成像、疾病诊断、肿瘤治疗、材料科学等领域得到了广泛的应用。PNPs的光散射光学性质可以通过调节其大小、组成、形貌和微环境来控制，可用于生化和药物分析。此外，由于单粒子散射显微技术具有高空间分辨率和高灵敏度，借助PNPs具有的独特局域表面等离子体共振(LSPR)特性，可在单颗粒水平进行实时成像。根据PNPs的大小、组成、形态、微环境或耦合变化引起的信号变化，研究人员发展了多种显微成像分析方法，主要分为4种，包括散射光谱的波长位移、单粒子散射强度的变化、高通量RGB分析和计数方法。基于纳米颗粒LSPR散射光谱位移变化的方法准确、灵敏，但需要昂贵的单颗粒散射光谱仪和复杂的操作。基于纳米颗粒散射强度变化的方法简单可行，但易受纳米颗粒粒径和曝光时间等因素的影响。高通量RGB分析方法灵敏度高、成本低，但不适用于颜色变化不明显的单颗粒分析，且重复性差。单粒子计数法灵敏度高，但有时粒子分布不均匀，背景杂质的干扰限制了方法的准确度。因此，这4种定量方法各有优缺点。此外，近年来逐渐发展了一些新的定量方法。例如，研究人员开发了新的时间分辨分析定量方法，并将暗场显微镜与偏振器、滤光片等光学器件相结合以消除背景干扰，以及与电化学、拉曼等仪器相结合以扩大应用范围。此外，为提高分析方法的准确度和灵敏度，暗场显微镜与深度学习、云计算、人工智能等现代计算机科学技术的结合，越来越受到人们的欢迎。基于以上原因，该文重点介绍了单粒子光散射显微镜在生化和药物分析领域的应用，总结了近年来的最新研究进展，讨论了单粒子光散射显微镜在定量分析中的几种主要定量方法，提出了未来的发展趋势，以期为相关研究领域的新人提供一定的学术参考。 |
| 中文关键词:等离子体纳米颗粒 暗场成像 单颗粒光谱 计数法 RGB颜色分析 |
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| Application of Single-particle Light Scattering Microscopic Technique in Biochemical and Medicine Analysis |
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| Abstract:Plasmonic nanoparticles(PNPs) have been widely used in the fields of biochemical sensing,bioimaging,disease diagnosis,cancer therapy and materials science,etc due to their advantages of small size,easy surface modification,good biocompatibility and low toxicity.The light-scattering optical properties of plasmonic nanoparticles could be controllably adjusted by tuning their size,composition,morphology and microenvironment,which could be developed for biochemical and pharmaceutical analysis.In addition,with the help of unique local surface plasmon resonance(LSPR) characteristics of plasmonic nanoparticles,a single-particle light scattering microscopy equiped with high spatial resolution and sensitivity could be used to obtain real-time image at a single particle level.Then,a variety of microscopic imaging analysis methods have been developed according to the signal variation resulting from changes of the size,composition,morphology,microenvironment or coupling,which could be mainly classified into four kinds of methods,including wavelength shift of scattering spectrum,change of single-particle scattering intensity,high-throughput RGB analysis and counting method.The method based on the change of wavelength shift of nanoparticles LSPR scattering spectra is accurate and sensitive,but it requires an expensive single-particle scattering spectrophotometer and a complex operation.The method based on the change of scattering intensity is simple and feasible,but it is easily affected by various factors such as the nanoparticles size and exposure time.High-throughput RGB analysis method has high accuracy and low cost,but it is not applicable for the analysis of single particle whose color change is not obvious,and whats more the reproducibility of this method is poor.The method of single particle counting is highly sensitive,but sometimes the particle will not distribute evenly and the interference coming from the background impurities limits the accuracy of the method.Therefore,these four quantitative methods have their own advantages and disadvantages.In addition,some new quantitative methods have been gradually developed in recent years.For example,the researchers have developed new quantitative methods for time-resolved analysis,combining the dark field microscopy with optical devices such as polarizers and filters to eliminate background interference,as well as instruments such as electrochemistry and Raman to expand the scope of application.Furthermore,in order to improve the accuracy and sensitivity of analytical methods,combinations of the dark-field microscopy with some modern computer science technologies,such as deep learning,cloud computing and artificial intelligence,have become more and more popular.Based on the above reasons,this mini review focuses on the application of single-particle light scattering microscopy in the field of biochemical and pharmaceutical analysis,then the recent new research progress is summarized,and several main quantitative methods of single-particle light scattering microscopy in quantitative analysis are discussed,and finally their future development trends are put forward.It is expected that the review could provide a certain academic reference for the new comer. |
| Key Words:plasmonic nanoparticles dark-field imaging single particle spectroscopy counting method RGB color analysis |
| 引用本文：程茹,李孟效,黄承志,王健.单颗粒光散射显微成像技术在生化医药分析中的应用[J].分析测试学报,2021,40(6):828-835. |
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