Abstract
Single-molecule localization microscopy (SMLM) enables super-resolution imaging on conventional fluorescent microscopes. Spectroscopic SMLM (sSMLM) further allows highly multiplexed super-resolution imaging. We report an easy-to-implement symmetrically dispersed dual-wedge prism (SDDWP)-sSMLM design that maximizes photon utilization. We first symmetrically dispersed photons to the −1st and +1st orders in an optical assembly using two identical dual-wedge prisms (DWPs). Then we computationally extracted the fluorophores’ spatial position and spectral characteristics using photons in both the −1st and +1st orders. Theoretical analysis and experimental validation showed lateral and spectral precisions of 10.1 nm and 0.3 nm, respectively, representing improvements of 28% and 48% over our previous DWP-based system, where emitted photons are divided separately for spatial and spectral analyses.
Citation
Wei-Hong Yeo, Benjamin Brenner, Youngseop Lee, Junghun Kweon, Cheng Sun, and Hao F. Zhang. 2024. “Maximizing photon utilization in spectroscopic single-molecule localization microscopy using symmetrically dispersed dual-wedge prisms.” BioRxiv. https://doi.org/10.1101/2024.05.12.593746.
@article{Yeo2024,
author = {Wei-Hong Yeo and Benjamin Brenner and Youngseop Lee and Junghun Kweon and Cheng Sun and Hao F. Zhang},
doi = {10.1101/2024.05.12.593746},
journal = {BioRxiv},
title = {Maximizing photon utilization in spectroscopic single-molecule localization microscopy using symmetrically dispersed dual-wedge prisms},
year = {2024}}