Janelia Research Campus, HHMI, Ashburn, VA, United States.
Methods Cell Biol. 2021;161:1-14. doi: 10.1016/bs.mcb.2020.04.015. Epub 2020 May 30.
Super-resolution microscopy methods circumvent the classical diffraction limit of optical microscopy using combinations of specially engineered excitation light, fluorescent dyes, highly sensitive detectors, and reconstruction algorithms. Protein-retention expansion microscopy (ExM) is a method to physically expand biological specimens, enabling effectively sub-diffraction limited imaging on standard microscopes with standard staining reagents. Specimen expansion is driven by a swellable gel material that can be synthesized in situ using off-the-shelf chemicals and materials. The expansion material and process are robust and amenable to further development, which has enabled the emergence of numerous ExM variants with extended capabilities from multiple independent labs. The method presented here is useful for routine expansion of tissue slices and adherent or floating cultured cells, and also forms the basis for these variant methods.
超分辨率显微镜方法通过组合特殊设计的激发光、荧光染料、高灵敏度探测器和重建算法来突破光学显微镜的经典衍射极限。蛋白保留膨胀显微镜（ExM）是一种物理膨胀生物样本的方法，可在具有标准染色试剂的标准显微镜上实现有效亚衍射极限成像。样本的膨胀是由可溶胀的凝胶材料驱动的，该材料可以使用现成的化学品和材料在现场合成。扩展材料和过程坚固且易于进一步开发，这使得来自多个独立实验室的具有扩展功能的许多 ExM 变体得以出现。这里介绍的方法可用于常规扩展组织切片以及贴壁或悬浮培养的细胞，也是这些变体方法的基础。
