Learning an optimal PSF-pair for ultra-dense 3D localization microscopy

Nehme, Elias; Ferdman, Boris; Weiss, Lucien E.; Naor, Tal; Freedman, Daniel; Michaeli, Tomer; Shechtman, Yoav

doi:10.48550/arxiv.2009.14303

Cited by 2 publications

(2 citation statements)

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“…7 Deep-learning methods have also been successfully applied to optimize paired PSFs to achieve precise 3D localization at high densities. 43 Similar machinelearning approaches with the single-spot RPSF such as that employed here can also potentially improve its error performance even under lower SNR and higher emitter-density conditions.…”

Section: Spp Localization Performancementioning

confidence: 95%

Three-dimensional tracking using a single-spot rotating point spread function created by a multiring spiral phase plate

et al. 2022

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Significance: Three-dimensional (3D) imaging and object tracking is critical for medical and biological research and can be achieved by multifocal imaging with diffractive optical elements (DOEs) converting depth (z) information into a modification of the two-dimensional image. Physical insight into DOE designs will spur this expanding field.Aim: To precisely track microscopic fluorescent objects in biological systems in 3D with a simple low-cost DOE system.Approach: We designed a multiring spiral phase plate (SPP) generating a single-spot rotating point spread function (SS-RPSF) in a microscope. Our simple, analytically transparent design process uses Bessel beams to avoid rotational ambiguities and achieve a significant depth range. The SPP was inserted into the Nomarski prism slider of a standard microscope. Performance was evaluated using fluorescent beads and in live cells expressing a fluorescent chromatin marker.Results: Bead localization precision was <25 nm in the transverse dimensions and ≤70 nm along the axial dimension over an axial range of 6 μm. Higher axial precision (≤50 nm) was achieved over a shallower focal depth of 2.9 μm. 3D diffusion constants of chromatin matched expected values.Conclusions: Precise 3D localization and tracking can be achieved with a SS-RPSF SPP in a standard microscope with minor modifications.

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Section: Spp Localization Performancementioning

confidence: 95%

Three-dimensional tracking using a single-spot rotating point spread function created by a multiring spiral phase plate

et al. 2022

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“…In addition, with the development of super-resolution microscopy techniques mentioned above, the method to improve precision of locating a single emitter is also important. Efforts along this line include designing optimal point spread functions (PSF) [41,42] and the quantumlimited longitudinal localisation of a single emitter [43].…”

Section: Introductionmentioning

confidence: 99%

Quantum-limited Localisation and Resolution in Three Dimensions

Wang,

Xu,

Zhang

2020

Preprint

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As a method to extract information from optical system, imaging can be viewed as a parameter estimation problem. The fundamental precision in locating one emitter or estimating the separation between two incoherent emitters is bounded below by the multiparameter quantum Cramér-Rao bound (QCRB). Multiparameter QCRB gives an intrinsic bound in parameter estimation. We determine the ultimate potential of quantum-limited imaging for improving the resolution of a farfield, diffraction-limited within the paraxial approximation. We show that the quantum Fisher information matrix (QFIm) about one emitter's position is independent on the true value of it. We calculate the QFIm of two unequal-brightness emitters' relative positions and intensities, the results show that only when the relative intensity and centroids of two point sources including longitudinal and transverse direction are known exactly, the separation in different directions can be estimated simultaneously with finite precision. Our results give the upper bounds on certain far-field imaging technology and will find wide applications from microscopy to astrometry.

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Learning an optimal PSF-pair for ultra-dense 3D localization microscopy

Cited by 2 publications

References 100 publications

Three-dimensional tracking using a single-spot rotating point spread function created by a multiring spiral phase plate

Three-dimensional tracking using a single-spot rotating point spread function created by a multiring spiral phase plate

Quantum-limited Localisation and Resolution in Three Dimensions

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