Universal inverse modeling of point spread functions for SMLM localization and microscope characterization.
Nat Methods
; 21(6): 1082-1093, 2024 Jun.
Article
in En
| MEDLINE
| ID: mdl-38831208
ABSTRACT
The point spread function (PSF) of a microscope describes the image of a point emitter. Knowing the accurate PSF model is essential for various imaging tasks, including single-molecule localization, aberration correction and deconvolution. Here we present universal inverse modeling of point spread functions (uiPSF), a toolbox to infer accurate PSF models from microscopy data, using either image stacks of fluorescent beads or directly images of blinking fluorophores, the raw data in single-molecule localization microscopy (SMLM). Our modular framework is applicable to a variety of microscope modalities and the PSF model incorporates system- or sample-specific characteristics, for example, the bead size, field- and depth- dependent aberrations, and transformations among channels. We demonstrate its application in single or multiple channels or large field-of-view SMLM systems, 4Pi-SMLM, and lattice light-sheet microscopes using either bead data or single-molecule blinking data.
Full text:
1
Database:
MEDLINE
Main subject:
Single Molecule Imaging
/
Microscopy, Fluorescence
Language:
En
Year:
2024
Type:
Article