瑞士洛桑联邦理工学院Suliana Manley、Dora Mahecic等研究人员合作开发出均匀多焦点激发技术用于高通量超分辨率成像。相关论文于2020年6月22日在线发表于《自然—方法学》。

研究人员报道了用于场非依赖成像的多焦点平面照明（mfFIFI）技术。通过将mfFIFI集成到即时结构照明显微镜（iSIM）中，研究人员将视场（FOV）扩展到100×100 μm²以上，同时保持高速、多色、体积成像，且为衍射极限分辨率的两倍。研究人员通过拼接相邻图像来进一步扩展有效FOV，从而对数十个细胞进行快速活细胞超分辨率成像。

最后，研究人员将平场iSIM与超微结构扩展显微镜相结合，以约35 nm的有效分辨率每小时对人类细胞中数百个中心体或成千上万个纯化的莱茵衣藻中心体进行了三维（3D）图像。这些大型数据集的分类和粒子平均可以对中心体微管的翻译后修饰进行3D映射，从而揭示其覆盖范围和位置的差异。

据介绍，超分辨率显微镜已成为生物学研究中的常用工具。但是，成像通量仍然是获取定量生物学所需大型数据集的主要瓶颈。

附：英文原文

Title: Homogeneous multifocal excitation for high-throughput super-resolution imaging

Author: Dora Mahecic, Davide Gambarotto, Kyle M. Douglass, Denis Fortun, Niccol Banterle, Khalid A. Ibrahim, Maeva Le Guennec, Pierre Gnczy, Virginie Hamel, Paul Guichard, Suliana Manley

Issue&Volume: 2020-06-22

Abstract: Super-resolution microscopies have become an established tool in biological research. However, imaging throughput remains a main bottleneck in acquiring large datasets required for quantitative biology. Here we describe multifocal flat illumination for field-independent imaging (mfFIFI). By integrating mfFIFI into an instant structured illumination microscope (iSIM), we extend the field of view (FOV) to >100×100μm2 while maintaining high-speed, multicolor, volumetric imaging at double the diffraction-limited resolution. We further extend the effective FOV by stitching adjacent images for fast live-cell super-resolution imaging of dozens of cells. Finally, we combine our flat-fielded iSIM with ultrastructure expansion microscopy to collect three-dimensional (3D) images of hundreds of centrioles in human cells, or thousands of purified Chlamydomonas reinhardtii centrioles, per hour at an effective resolution of ~35nm. Classification and particle averaging of these large datasets enables 3D mapping of posttranslational modifications of centriolar microtubules, revealing differences in their coverage and positioning.

DOI: 10.1038/s41592-020-0859-z

Source: https://www.nature.com/articles/s41592-020-0859-z