Multicolor super-resolution imaging using spectroscopic single-molecule localization microscopy with optimal spectral dispersion

Yang Zhang, Ki Hee Song, Biqin Dong, Janel L. Davis, Guangbin Shao, Cheng Sun, Hao F. Zhang*

*Corresponding author for this work

Research output: Contribution to journalArticle

6 Scopus citations


We developed transmission diffraction grating-based spectroscopic single-molecule localization microscopy (sSMLM) to collect the spatial and spectral information of single-molecule blinking events concurrently. We characterized the spectral heterogeneities of multiple far-red emitting dyes in a high-throughput manner using sSMLM. We also investigated the influence of spectral dispersion on the single-molecule identification performance of fluorophores with large spectral overlapping. The careful tuning of spectral dispersion in grating-based sSMLM permitted simultaneous three-color super-resolution imaging in fixed cells with a single objective lens at a relatively low photon budget. Our sSMLM has a compact optical design and can be integrated with conventional localization microscopy to provide add-on spectroscopic analysis capability.

Original languageEnglish (US)
Pages (from-to)2248-2255
Number of pages8
JournalApplied optics
Issue number9
StatePublished - Jan 1 2019


ASJC Scopus subject areas

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

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