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Sparse deconvolution improves the resolution of live-cell super-resolution fluorescence microscopy

  • Weisong Zhao
  • , Shiqun Zhao
  • , Liuju Li
  • , Xiaoshuai Huang
  • , Shijia Xing
  • , Yulin Zhang
  • , Guohua Qiu
  • , Zhenqian Han
  • , Yingxu Shang
  • , De en Sun
  • , Chunyan Shan
  • , Runlong Wu
  • , Lusheng Gu
  • , Shuwen Zhang
  • , Riwang Chen
  • , Jian Xiao
  • , Yanquan Mo
  • , Jianyong Wang
  • , Wei Ji
  • , Xing Chen
  • Baoquan Ding, Yanmei Liu, Heng Mao, Bao Liang Song, Jiubin Tan, Jian Liu, Haoyu Li*, Liangyi Chen*
*Corresponding author for this work
  • Harbin Institute of Technology
  • Peking University
  • National Center for Nanoscience and Technology
  • CAS - Institute of Biophysics
  • Wuhan University
  • South China Normal University
  • Harbin Institute of Technology
  • Beijing Academy of Artificial Intelligence

Research output: Contribution to journalArticlepeer-review

Abstract

A main determinant of the spatial resolution of live-cell super-resolution (SR) microscopes is the maximum photon flux that can be collected. To further increase the effective resolution for a given photon flux, we take advantage of a priori knowledge about the sparsity and continuity of biological structures to develop a deconvolution algorithm that increases the resolution of SR microscopes nearly twofold. Our method, sparse structured illumination microscopy (Sparse-SIM), achieves ~60-nm resolution at a frame rate of up to 564 Hz, allowing it to resolve intricate structures, including small vesicular fusion pores, ring-shaped nuclear pores formed by nucleoporins and relative movements of inner and outer mitochondrial membranes in live cells. Sparse deconvolution can also be used to increase the three-dimensional resolution of spinning-disc confocal-based SIM, even at low signal-to-noise ratios, which allows four-color, three-dimensional live-cell SR imaging at ~90-nm resolution. Overall, sparse deconvolution will be useful to increase the spatiotemporal resolution of live-cell fluorescence microscopy.

Original languageEnglish
Pages (from-to)606-617
Number of pages12
JournalNature Biotechnology
Volume40
Issue number4
DOIs
StatePublished - Apr 2022

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