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光热显微术是近年来获得广泛关注和长足发展的一种新型光学显微成像技术,能够实现单个纳米粒子甚至单分子的免标记光学成像。其成像原理是利用先进的光学方法探测单分子或单纳米粒子吸收特定波长激发光后所产生的局域温度和介质折射率的微小变化,从而定量研究观测对象的光热特性。由于无辐射弛豫是激发态分子回到基态的优势过程,分子的光热特性相比于荧光特性更具有普遍意义。凭借无需标记、高灵敏度和信号稳定等优点,近十年来,关于单分子和单纳米粒子的光热显微成像研究不断取得突破,并在纳米科学和生命科学等领域获得越来越多的发展和应用,展现出了蓬勃的生命力和良好的发展前景。本文重点综述了光热显微技术的成像原理、发展历程、技术特色以及系统优化方法,列举了光热成像在活细胞研究和生物学领域的应用,最后总结了光热成像的优缺点并分析其主要面临的挑战以及未来的发展趋势,希望吸引更多的研究人员加入到这一新技术的研究队伍中来。
Photothermal microscopy is a new type of optical microscopy technique that has gained much attention and developed rapidly in recent years. It can realize single label-free optical imaging of single nanoparticle or even single molecule. The imaging principle is the use of advanced optical methods to detect the single-molecule or single-nanoparticle absorption of specific wavelength excitation light generated after the local temperature and medium refractive index small changes in order to quantitatively study the observed object’s photothermal properties. Since the non-radiative relaxation is the dominant process of the excited state molecules returning to the ground state, the photothermal properties of the molecules have more universal significance than the fluorescence properties. With the advantages of no marking, high sensitivity and signal stability, breakthroughs have been made in the research of photothermal microscopy imaging of single molecules and single nanoparticles in recent ten years, and more and more developments have been made in the fields of nanoscience and life science And application, has shown a vibrant vitality and good prospects for development. In this paper, the imaging principles, development history, technical characteristics and system optimization methods of photothermal microscopy are reviewed. The application of photothermal imaging in the field of living cells and biology is listed. Finally, the advantages and disadvantages of photothermal imaging are summarized and analyzed Its main challenges and the future development trend, hoping to attract more researchers to join the research team of this new technology.