Mechanical-scan-free multicolor super-resolution imaging with diffractive spot array illumination

Ning Xu, Sarah E. Bohndiek, Zexing Li, Cilong Zhang and Qiaofeng Tan ()
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Ning Xu: Tsinghua University
Sarah E. Bohndiek: University of Cambridge
Zexing Li: University of Cambridge
Cilong Zhang: Tsinghua University
Qiaofeng Tan: Tsinghua University

Nature Communications, 2024, vol. 15, issue 1, 1-11

Abstract: Abstract Point-scanning microscopy approaches are transforming super-resolution imaging. Despite achieving parallel high-speed imaging using multifocal techniques, efficient multicolor imaging methods with high-quality illumination are currently lacking. In this paper, we present for the first time Mechanical-scan-free multiColor Super-resolution Microscopy (MCoSM) with spot array illumination, which enables mechanical-scan-free super-resolution imaging with adjustable resolution and a good effective field-of-view based on spatial light modulators. Through 100–2,500 s super-resolution spot illumination with different effective fields of view for imaging, we demonstrate the adjustable capacity of MCoSM. MCoSM extends existing spectral imaging capabilities through a time-sharing process involving different color illumination with phase-shift scanning while retaining the spatial flexibility of super-resolution imaging with diffractive spot array illumination. To demonstrate the prospects of MCoSM, we perform four-color imaging of fluorescent beads at high resolution. MCoSM provides a versatile platform for studying molecular interactions in complex samples at the nanoscale level.

Date: 2024
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DOI: 10.1038/s41467-024-48482-z

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