Journal
APPLIED OPTICS
Volume 58, Issue 9, Pages 2248-2255Publisher
OPTICAL SOC AMER
DOI: 10.1364/AO.58.002248
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Funding
- National Science Foundation (NSF) [CBET-1706642, EEC-1530734, EFRI-1830969]
- National Institutes of Health (NIH) [R01EY026078, R01EY029121]
- Northwestern University Innovative Initiative Incubator (I3) Award
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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. (C) 2019 Optical Society of America
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