Open AccessMulticolor super-resolution imaging using spectroscopic single-molecule localization microscopy with optimal spectral dispersion
Author(s) -
Yang Zhang,
Ki-Hee Song,
Biqin Dong,
Janel L. Davis,
Guangbin Shao,
Cheng Sun,
Hao F. Zhang
Publication year - 2019
Publication title -
applied optics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.668
H-Index - 197
eISSN - 2155-3165
pISSN - 1559-128X
DOI - 10.1364/ao.58.002248
Subject(s) - optics , microscopy , spectral resolution , materials science , grating , dispersion (optics) , resolution (logic) , spectral imaging , diffraction grating , physics , spectral line , computer science , astronomy , artificial intelligence
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.