Improving spatial precision and field-of-view in wavelength-tagged single-particle tracking using spectroscopic single-molecule localization microscopy

Benjamin Brenner, Ki Hee Song, Cheng Sun, Hao F. Zhang*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Spectroscopic single-molecule localization microscopy (sSMLM) generates super-resolution images of single molecules while simultaneously capturing the spectra of their fluorescence emissions. However, sSMLM splits photons from single-molecule emissions into a spatial channel and a spectral channel, reducing both channels’ precisions. It is also challenging in transmission grating-based sSMLM to achieve a large field-of-view (FOV) and avoid overlap between the spatial and spectral channels. The challenge in FOV has further significance in single-molecule tracking applications. In this work, we analyzed the correlation between the spatial and spectral channels in sSMLM to improve its spatial precision, and we developed a split-mirror assembly to enlarge its FOV. We demonstrate the benefits of these improvements by tracking quantum dots. We also show that we can reduce particle-identification ambiguity by tagging each particle with its unique spectral characteristics.

Original languageEnglish (US)
Pages (from-to)3647-3658
Number of pages12
JournalApplied optics
Volume60
Issue number13
DOIs
StatePublished - May 1 2021

ASJC Scopus subject areas

  • Atomic and Molecular Physics, and Optics
  • Engineering (miscellaneous)
  • Electrical and Electronic Engineering

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