一种改良的放大分析蛋白质组学（MAP）方法，用于超分辨率细胞成像，同时保留荧光。
A Modified Magnified Analysis of Proteome (MAP) Method for Super-Resolution Cell Imaging that Retains Fluorescence.
机构信息
The Spine and Spinal Cord Institute, Department of Neurosurgery, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, 06273, Republic of Korea.
Brain Korea 21 PLUS Project for Medical Science, Yonsei University, Seoul, 03722, Republic of Korea.
出版信息
Sci Rep. 2020 Mar 6;10(1):4186. doi: 10.1038/s41598-020-61156-2.
Biological systems consist of a variety of distinct cell types that form functional networks. Super-resolution imaging of individual cells is required for better understanding of these complex systems. Direct visualization of 3D subcellular and nano-scale structures in cells is helpful for the interpretation of biological interactions and system-level responses. Here we introduce a modified magnified analysis of proteome (MAP) method for cell super-resolution imaging (Cell-MAP) which preserves cell fluorescence. Cell-MAP expands cells more than four-fold while preserving their overall architecture and three-dimensional proteome organization after hydrogel embedding. In addition, Optimized-Cell-MAP completely preserves fluorescence and successfully allows for the observation of tagged small molecular probes containing peptides and microRNAs. Optimized-Cell-MAP further successfully applies to the study of structural characteristics and the identification of small molecules and organelles in mammalian cells. These results may give rise to many other applications related to the structural and molecular analysis of smaller assembled biological systems.
生物系统由多种不同的细胞类型组成，这些细胞类型形成功能网络。为了更好地理解这些复杂的系统，需要对单个细胞进行超分辨率成像。直接观察细胞内的三维亚细胞和纳米级结构有助于解释生物相互作用和系统水平的反应。在这里，我们引入了一种改良的放大分析蛋白质组学（MAP）方法，用于细胞超分辨率成像（Cell-MAP），该方法可以保留细胞荧光。Cell-MAP 在水凝胶嵌入后可以将细胞放大四倍以上，同时保留其整体结构和三维蛋白质组组织。此外，优化后的 Cell-MAP 可以完全保留荧光，并成功地允许观察含有肽和 microRNA 的标记小分子探针。优化后的 Cell-MAP 进一步成功地应用于研究哺乳动物细胞中的结构特征和小分子及细胞器的鉴定。这些结果可能会引发许多与较小的组装生物系统的结构和分子分析相关的其他应用。
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