Near-field optics-based scanning microsphere lens super-resolution microscope and imaging method thereof

Abstract

The invention discloses a scanning microsphere lens super-resolution microscope based on near-field optics used for a liquid environment. The scanning microsphere lens super-resolution microscope includes an optical microscope, a microsphere lens, a container, a glass tube probe, a sampling electrode, a grounding electrode, a DC power supply, current amplifier, a real-time feedback and scanning controller, and a pre-adjustment displacement table and a scanning table that can move in three-dimensional directions, a positive electrode of the DC power supply and the sampling electrode are connected with a corresponding end of the current amplifier separately, the glass tube probe includes a straight section connected with a pre-adjustment displacement table and a slant section connected withthe straight section, the bottom end of the slant section is provided with a needle tip part, the sampling electrode is arranged in the glass tube probe, the microsphere lens is arranged in the needle tip part of the slant section, and the vertex of the microsphere lens is almost the same position as the bottom of the microsphere lens. The scanning microsphere lens super-resolution microscope based on near-field optics is mainly used in the fields of life science and the like which require nanoscale super-resolution real-time dynamic observation and operation in a liquid environment.

Description

technical field [0001] 本发明涉及到一种基于扫描离子电导显微镜和微球透镜的光学超分辨率大范围扫描成像系统,主要用于液态环境中需要纳米级超分辨率实时动态观测和操作的生命科学等领域。 Background technique [0002] 十九世纪末,德国科学家恩斯特·阿贝定义了光学显微镜的分辨率,认为是光波长的一半,即:约为0.2微米,这就是著名的光学衍射极限。因此,在二十世纪的大部分时间里,科学家们都相信光学显微镜永远无法突破衍射极限的限制。然而,随着科学研究的深入,尤其是生物科学领域的研究,人们早已开始研究细胞的内部结构,因此需要对细胞内部的组织结构进行活体观测。电子显微镜只能观测死亡细胞的表面,无法观测活体细胞的内部结构。前不久,研究者发现微球透镜可实现超分辨率的光学成像,技术研究成果最早发表于2009 年Nature杂志上(www.nature.com / doifinder / 10.1038 / nature08173),当时的分辨率为220nm。两年后,这个分辨率被提高到了50nm(Nature Communication,DOI:10.1038 / ncomms1211)。2012年,研究者又发现,不但微球透镜的超分辨率成像可以在空气中实现,而且还可以在液体中实现,并能达到与空气中成像相同的分辨率(Appl.Phys.Lett.101,141128;https: / / doi.org / 10.1063 / 1.4757600)。不同于荧光显微镜的是,基于微球透镜的纳米成像不需要荧光染剂,而且是在自然光照条件下,适合任何样品成像,因而更具有广泛应用的意义。 [0003] 目前,通过与原子力显微镜的反馈控制机理相结合,使微球透镜与样品品表面的距离获得精确控制,从而实现对样品表面进行大范围动态扫描超分辨率成像(NatureCommunication,DOI:10.1038 / ncomms13748)。但是,此类反馈机制只适宜在空气中成像,且系统本身光路复杂,造价非常昂贵,不能在液态环境中应用。由于在液态环境中缺乏有效的反馈控制手段精确控制微球透镜和样品表面的距离,目前液体中基于微球透镜的超分辨率成像系统,一般只能在固定位置成像,不具备任意位置和大范围扫描成像能力。 Contents of the invention [0004...

AI Technical Summary

Problems solved by technology

However, this kind of feedback mechanism is only suitable for imaging in air, and the optical path of the system itself is complex, the cost is very expensive, and it cannot be applied in a liquid environment

Due to the lack of effective feedback control means to accurately control the distance between the microsphere lens and the sample surface in the liquid environment, the current super-resolution imaging system based on the microsphere lens in the liquid can only image at a fixed position, and does not have any position and large size. Range Scan Imaging Capabilities

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