A statistical resolution measure of fluorescence microscopy with finite photons

02/07/2023

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by   Yilun Li, et al.

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First discovered by Ernest Abbe in 1873, the resolution limit of a far-field microscope is considered determined by the numerical aperture and wavelength of light, approximately λ/2NA. With the advent of modern fluorescence microscopy and nanoscopy methods over the last century, it is recognized that Abbe's resolution definition alone could not solely characterize the resolving power of the microscope system. To determine the practical resolution of a fluorescence microscope, photon noise remains one essential factor yet to be incorporated in a statistics-based theoretical framework. Techniques such as confocal allow trading photon noise in gaining its resolution limit, which may increase or worsen the resolvability towards fluorescently tagged targets. Proposed as a theoretical measure of fluorescence microscopes' resolving power with finite photons, we quantify the resolvability of periodic structures in fluorescence microscopy systems considering both the diffraction limit and photon statistics. Using the Cramer-Rao Lower Bound of a parametric target, the resulting precision lower bound establishes a practical measure of the theoretical resolving power for various modern fluorescence microscopy methods in the presence of noise.