Maximizing photon utilization in spectroscopic single-molecule localization microscopy using symmetrically dispersed dual-wedge prisms.

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 -1^st and +1^st 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 -1^st and +1^st orders. Theoretical analysis and experimental validation showed lateral and spectral precisions of 10.1 nm and 0.3 nm, respectively, representin

SUBMITTER: Yeo WH 

PROVIDER: S-EPMC11118294 | biostudies-literature | 2024 May

REPOSITORIES: biostudies-literature