均勻多焦點激發技術可用於高通量超解析度成像
作者：
小柯機器人發布時間：2020/6/25 13:23:19
瑞士洛桑聯邦理工學院Suliana Manley、Dora Mahecic等研究人員合作開發出均勻多焦點激發技術用於高通量超解析度成像。相關論文於2020年6月22日在線發表於《自然—方法學》。
研究人員報導了用於場非依賴成像的多焦點平面照明（mfFIFI）技術。通過將mfFIFI集成到即時結構照明顯微鏡（iSIM）中，研究人員將視場（FOV）擴展到100×100 μm2以上，同時保持高速、多色、體積成像，且為衍射極限解析度的兩倍。研究人員通過拼接相鄰圖像來進一步擴展有效FOV，從而對數十個細胞進行快速活細胞超解析度成像。
最後，研究人員將平場iSIM與超微結構擴展顯微鏡相結合，以約35 nm的有效解析度每小時對人類細胞中數百個中心體或成千上萬個純化的萊茵衣藻中心體進行了三維（3D）圖像。這些大型數據集的分類和粒子平均可以對中心體微管的翻譯後修飾進行3D映射，從而揭示其覆蓋範圍和位置的差異。
據介紹，超解析度顯微鏡已成為生物學研究中的常用工具。但是，成像通量仍然是獲取定量生物學所需大型數據集的主要瓶頸。
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