Symmetrically dispersed spectroscopic single-molecule localization microscopy

Ki-Hee Song; Yang Zhang; Benjamin Brenner; Cheng Sun; Hao F. Zhang

doi:10.1038/s41377-020-0333-9

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Symmetrically dispersed spectroscopic single-molecule localization microscopy

Original Articles

- Symmetrically dispersed spectroscopic single-molecule localization microscopy
- Light: Science & Applications Vol. 9, Issue 4, Pages: 981-990(2020)
- Affiliations：
  
  1.Department of Biomedical Engineering, Northwestern University, 2145 Sheridan Rd., Evanston, IL 60208, USA
  2.Department of Mechanical Engineering, Northwestern University, 2145 Sheridan Rd., Evanston, IL 60208, USA
- Author bio：
  
  Hao F. Zhang (hfzhang@northwestern.edu)
- Funds：
- DOI：10.1038/s41377-020-0333-9
  CLC：
- Published：2020，
  
  Published Online：25 May 2020，
  
  Received：20 January 2020，
  
  Revised：02 May 2020，
  
  Accepted：11 May 2020
- Accepted：
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Song, K. H. et al. Symmetrically dispersed spectroscopic singlemolecule localization microscopy. Light: Science & Applications, 9, 981-990 (2020).
DOI：

Song, K. H. et al. Symmetrically dispersed spectroscopic singlemolecule localization microscopy. Light: Science & Applications, 9, 981-990 (2020). DOI： 10.1038/s41377-020-0333-9.

摘要

Abstract

Spectroscopic single-molecule localization microscopy (sSMLM) was used to achieve simultaneous imaging and spectral analysis of single molecules for the first time. Current sSMLM fundamentally suffers from a reduced photon budget because the photons from individual stochastic emissions are divided into spatial and spectral channels. Therefore

both spatial localization and spectral analysis only use a portion of the total photons

leading to reduced precisions in both channels. To improve the spatial and spectral precisions

we present symmetrically dispersed sSMLM

or SDsSMLM

to fully utilize all photons from individual stochastic emissions in both spatial and spectral channels. SDsSMLM achieved 10-nm spatial and 0.8-nm spectral precisions at a total photon budget of 1000. Compared with the existing sSMLM using a 1:3 splitting ratio between spatial and spectral channels

SDsSMLM improved the spatial and spectral precisions by 42% and 10%

respectively

under the same photon budget. We also demonstrated multicolour imaging of fixed cells and three-dimensional single-particle tracking using SDsSMLM. SDsSMLM enables more precise spectroscopic single-molecule analysis in broader cell biology and material science applications.

关键词

Keywords

references

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