Enhanced 4Pi single-molecule localization microscopy with coherent pupil based localization
Abstract
Over the last decades, super-resolution techniques have revolutionized the field of fluorescence microscopy. Among them, interferometric or 4Pi microscopy methods exhibit supreme resolving power in the axial dimension. Combined with single-molecule detection/localization and adaptive optics, current 4Pi microscopy methods enabled 10–15 nm isotropic 3D resolution throughout whole cells. However, further improving the achieved 3D resolution poses challenges arising from the complexity of single-molecule emission patterns generated by these coherent single-molecule imaging systems. These complex emission patterns render a large portion of information carrying photons unusable. Here, we introduce a localization algorithm that achieves the theoretical precision limit for a 4Pi based single-molecule switching nanoscopy (4Pi-SMSN) system, and demonstrate improvements in localization precision, accuracy as well as stability comparing with state-of-the-art 4Pi-SMSN methods.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- May 08, 2020
- Source ID
- 10.1038/s42003-020-0908-2
Entities
People
- Fang Huang
- Sheng Liu
Organizations
- National Institute of General Medical Sciences
- United States Department of Defense