Abstract
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 language | English (US) |
|---|---|
| Pages (from-to) | 2248-2255 |
| Number of pages | 8 |
| Journal | Applied optics |
| Volume | 58 |
| Issue number | 9 |
| DOIs | |
| State | Published - 2019 |
Funding
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. Acknowledgment. J. L. Davis thanks the support of NSF Graduate Research Fellowship 1000231682. C. Sun and H. F. Zhang have financial interests in Opticent Inc., which did not support this work. Other authors declare that there are no conflicts of interests related to this paper. National Science Foundation (NSF) (CBET-1706642, EEC-1530734, EFRI-1830969); National Institutes of Health (NIH) (R01EY026078, R01EY029121); Northwestern University Innovative Initiative Incubator (I3) Award.
ASJC Scopus subject areas
- Atomic and Molecular Physics, and Optics
- Engineering (miscellaneous)
- Electrical and Electronic Engineering