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Tuning axial and lateral localization precision in 3D super-resolution microscopy with variable astigmatism.

ABSTRACT: Astigmatism imaging is a three-dimensional (3D) single molecule fluorescence microscopy approach that yields super-resolved spatial information on a rapid time scale from a single image. It is ideally suited for resolving structures on a sub-micrometer scale and temporal behavior in the millisecond regime. While traditional astigmatism imaging utilizes a cylindrical lens, adaptive optics enables the astigmatism to be tuned for the experiment. We demonstrate here how the precisions in x, y, and z are inter-linked and vary with the astigmatism, z-position, and photon level. This experimentally driven and verified approach provides a guide for astigmatism selection in biological imaging strategies.

SUBMITTER: Chowdhury R

PROVIDER: S-EPMC10332797 | biostudies-literature | 2022 Nov

REPOSITORIES: biostudies-literature

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Tuning axial and lateral localization precision in 3D super-resolution microscopy with variable astigmatism.

Chowdhury Rajdeep R Sau Abhishek A Chao Jerry J Sharma Ankith A Musser Siegfried M SM

Optics letters 20221101 21

Astigmatism imaging is a three-dimensional (3D) single molecule fluorescence microscopy approach that yields super-resolved spatial information on a rapid time scale from a single image. It is ideally suited for resolving structures on a sub-micrometer scale and temporal behavior in the millisecond regime. While traditional astigmatism imaging utilizes a cylindrical lens, adaptive optics enables the astigmatism to be tuned for the experiment. We demonstrate here how the precisions in x, y, and z ...[more]

PMID: 37219314

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