Abstract

Many biological studies require the analysis of ultrastructural changes at the level of cell organelles and macromolecules. Since the resolution of modern confocal microscope is limited by the diffraction limit (200–300 nm), it is impossible to study such small objects using standard fluorescence microscopy. Ultra-high resolution microscopy methods require expensive equipment and are technically difficult in use, which in turn limits their widespread practical application. However, recently appeared methods make it possible to increase the resolution of microscopy not by improving the image registration system, but by physically isotropic expansion of a biological sample using a controlled chemical process. Due to this method, called expansion or expansive microscopy (ExM), it became possible to obtain three-dimensional images of samples with a resolution sufficient to study individual cell organelles using a conventional confocal microscope. This review covers the history of this method, its basic principles and examples of use in various fields of biology and medicine, as well as reflects future directions for improving this technology. The article discusses the methodological features of the ExM application in a study of brain tissue samples using the algorithm that allows adaptation of the standard protocol to the goals and objectives of a particular study.

中文翻译：

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膨胀显微镜法在神经生物学中的应用

摘要

许多生物学研究需要分析细胞器和大分子水平的超微结构变化。由于现代共聚焦显微镜的分辨率受到衍射极限（200-300 nm）的限制，因此不可能使用标准荧光显微镜研究如此小的物体。超高分辨率显微方法需要昂贵的设备并且在技术上使用困难，这反过来限制了其广泛的实际应用。然而，最近出现的方法不是通过改进图像配准系统，而是通过使用受控化学过程对生物样品进行物理各向同性膨胀来提高显微镜的分辨率。由于这种方法，称为膨胀或膨胀显微镜（ExM），获得具有足够分辨率的样品的 3D 图像成为可能，以便使用传统的共聚焦显微镜研究单个细胞器。这篇综述涵盖了该方法的历史、其基本原理和在生物学和医学各个领域的使用示例，并反映了改进该技术的未来方向。本文讨论了 ExM 应用程序在脑组织样本研究中的方法学特征，该算法使用的算法允许根据特定研究的目标和目的调整标准协议。并反映了改进这项技术的未来方向。本文讨论了 ExM 应用程序在脑组织样本研究中的方法学特征，该算法使用的算法允许根据特定研究的目标和目的调整标准协议。并反映了改进这项技术的未来方向。本文讨论了 ExM 应用程序在脑组织样本研究中的方法学特征，该算法使用的算法允许根据特定研究的目标和目的调整标准协议。