Department of Physics, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, Republic of Korea.
Graduate School of Nanoscience and Technology, KAIST, Daejeon 34141, Republic of Korea.
Nano Lett. 2022 Mar 23;22(6):2194-2201. doi: 10.1021/acs.nanolett.1c03691. Epub 2022 Mar 3.
Stochastic optical fluctuation imaging (SOFI) generates super-resolution fluorescence images by emphasizing the positions of fluorescent emitters via statistical analysis of their on-and-off blinking dynamics. In SOFI with speckle illumination (S-SOFI), the diffraction-limited grain size of the far-field speckles prevents independent blinking of closely located emitters, becoming a hurdle to realize the full super-resolution granted by SOFI processing. Here, we present a surface-sensitive super-resolution technique exploiting dynamic near-field speckle illumination to bring forth the full super-resolving power of SOFI without blinking fluorophores. With our near-field S-SOFI technique, up to 2.8- and 2.3-fold enhancements in lateral spatial resolution are demonstrated with computational and experimental fluorescent test targets labeled with conventional fluorophores, respectively. Fluorescent beads separated by 175 nm are also super-resolved by near-field speckles of 150 nm grain size, promising sub-100 nm resolution with speckle patterns of much smaller grain size.
随机光学波动成像（SOFI）通过对荧光发射器的开和关闪烁动力学进行统计分析，强调其位置，从而生成超分辨率荧光图像。在具有散斑照明的 SOFI（S-SOFI）中，远场散斑的衍射极限粒度限制了近距离发射体的独立闪烁，成为实现 SOFI 处理所赋予的全超分辨率的障碍。在这里，我们提出了一种利用动态近场散斑照明的表面敏感超分辨率技术，在不闪烁荧光团的情况下发挥 SOFI 的全超分辨率能力。通过我们的近场 S-SOFI 技术，分别使用计算和实验荧光测试目标对具有常规荧光团的标记进行了实验，证明了横向空间分辨率提高了 2.8 倍和 2.3 倍。通过 150nm 粒度的近场散斑还可以实现 175nm 间隔的荧光珠的超分辨，有望在更小粒度的散斑图案下实现小于 100nm 的分辨率。
