近年来，基于单分子定位的超分辨成像技术（也称为超分辨定位成像，包括PALM， STORM，dSTORM等）将荧光显微成像的空间分辨率提高到约20nm，使得生命科学领域的研 究人员能够从分子水平观测生物组织的精细结构和功能。dSTORM(direct Stochastic Optical Reconstruction Microscopy)是一种代表性的超分辨定位成像技术，该技术巧 妙地利用缓冲液来进行密集排列荧光分子的空间分离，进而利用单分子成像和定位来实 现超分辨成像。但是，因为长时间成像造成的缓冲液失效以及多色成像中不同荧光探针的 最佳缓冲液匹配问题，需要在dSTORM成像过程中及时更换缓冲液，而现有的成像装置无法 很好地实现缓冲液的更换。因此，本文设计制作了适用于dSOTRM成像系统的样品池，可以 在成像过程中方便地更换缓冲液，我们通过成像实验，初步证实该样品池可以应用于 dSTORM成像。77746

关键词 超分辨定位成像 缓冲液 样品池 成像实验

毕 业 设 计 说 明 书 外 文 摘 要

Title Design of Sample Chamber for Super-resolution Localization

Microscopy

Abstract

Recently, single-molecule localization-based super-resolution microscopy (also called super-resolution localization microscopy, including PALM, STORM, dSTORM etc) has pushed the spatial resolution of fluorescence microscopy to approximately 20 nm, and thus has been a significant tool in various biology application scenarios to reveal the working mechanisms of cellular processes at molecular level。 Direct stochastic optical reconstruction microscopy (dSTORM) is a representative super-resolution microscopy technique, which uses cleverly imaging buffers to spatially separate closely-packed fluorescence probes, and then takes advantages of single molecule imaging and localization to realize super-resolution imaging。 However, during the imaging progress in dSTORM, the imaging buffer needs to be replaced for either long-term imaging (due to buffer degradation) or multi-color imaging (because imaging buffer should be optimized for different flurosecent probes)。 But, it is difficult to replace the imaging buffers in current dSTORM systems。 In this paper, we design a sample chamber which is capable of providing ease replacement of imaging buffers during imaging。 Through dSTORM experiments, we verify that this sample chamber is applicable to dSTORM imaging。

Keywords super-resolution localization microscopy, buffer solution,   sample chamber, imaging experiment

本科毕业设计说明书 第 I 页

目 次

1 绪论 1

1。1 衍射极限 1

1。2 超分辨显微成像技术 2

1。3 成像缓冲液的更换 4

1。4 本文的主要内容 5

2 样品池的设计制作 6

2。1 样品池设计的初步探索 6

2。2 样品池的改进 9

2。3 本章小结