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 language | English (US) |
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Pages (from-to) | 3647-3658 |
Number of pages | 12 |
Journal | Applied optics |
Volume | 60 |
Issue number | 13 |
DOIs | |
State | Published - May 1 2021 |
Funding
Funding. National Science Foundation (CBET-1706642, EFMA-1830969); National Institutes of Health (R01EY019949, R01EY026078, R01GM139151, T32GM105538); Christine Enroth-Cugell and David Cugell Graduate Fellowship in Biomedical Engineering and Visual Neuroscience at Northwestern University.
ASJC Scopus subject areas
- Atomic and Molecular Physics, and Optics
- Engineering (miscellaneous)
- Electrical and Electronic Engineering