基于受激发射损耗显微术的活细胞和活体超分辨成像

Live Cell and In vivo Super-resolution Imaging Based on STED

细胞作为生命体基本的结构和功能单元,在生物、医学等领域有着非常重要的研究意义。随着现代科学和技术的发展,科学家们借助电镜对细胞以及细胞器的空间结构已经有非常清晰的认识,但是对它们的功能以及细胞之间的相互作用却了解得非常少,而这恰恰又是疾病治疗和药物开发亟需了解的信息,因此对离体活细胞(简称活细胞)和活体生物组织细胞(简称活体细胞)中亚细胞器的研究变得非常重要。然而细胞中许多细胞器的结构在纳米量级,传统的光学成像技术由于受到光学衍射极限的限制是无法观察到纳米量级的生物结构,因此光学超分辨成像技术是目前研究亚细胞器结构和功能的有效工具。在所有光学超分辨显微技术中,受激发射损耗显微术(stimulated emission depletionmicroscopy,STED)由于具有实时成像、三维超分辨和断层成像的能力,非常适合用于纳米尺度的活细胞和活体细胞成像研究,而且STED超分辨成像技术经过近几十年的发展,已经广泛用于活细胞甚至活体小鼠细胞的超分辨动态观测。本文总结了近年来活细胞和活体小鼠神经元细胞等领域STED超分辨成像的研究进展,介绍了用于活细胞和活体细胞STED超分辨成像的荧光染料和荧光蛋白的发展现状,讨论当前活细胞和活体细胞的超分辨成像面临的问题,以及对未来发展的展望。

As the basic structural and functional unit of life,cells have very important research significance in biology,medicine and other fields.With the development of modern science and technology,scientists have a very clear understanding of the spatial structure of cells and organelles with the help of electron microscopes.However,very little is known about their functions and the interactions between cells,and this is precisely the information that disease treatment and drug development urgently need to know.Therefore,the study of subcellular organelles in vitro living cells and in vivo living cells have become very important.However,the structure of many organelles in living cells are at the nanoscale.Traditional optical imaging techniques cannot observe nanoscale biological structures due to the limitation of the optical diffraction limit.Therefore,optical super-resolution imaging technology is an effective tool to study the structure and function of subcellular organelles.Among all optical super-resolution microscopy techniques,stimulated emission depletion(STED)super-resolution imaging technology has the capabilities of real-time,three-dimensional super-resolution and tomographic imaging.Therefore,the STED is very suitable for nanoscale live cell and vivo imaging studies.Moreover,STED superresolution imaging technology has been widely used for super-resolution dynamic observation of living cells and even living mouse cells after decades of development.This paper summarized the research progress of STED super-resolution imaging in the fields of in vitro living cells and in vivo mouse neurons in recent years,and introduces the development status of fluorescent dyes and fluorescent proteins for STED super-resolution imaging of in vitro and in vivo living cells.In vivo cells super-resolution imaging is very meaningful for understanding the nature of cells,but it has been more than 20 years since the STED super-resolution imaging technology was proposed to the present,and there are still very few literatures on in vivo cells super-resolution imaging.The main problem is that very few probes are available for in vivo cells super-resolution imaging.At the same time,the depth of in vivo cells super-resolution imaging is also very limited,mainly due to the lack of fluorescent probes in the infrared band.Finally,the future application prospects of in vivo cells super-resolution imaging are prospected.