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Resolution enhancement with deblurring by pixel reassignment (DPR).

ABSTRACT: Improving the spatial resolution of a fluorescence microscope has been an ongoing challenge in the imaging community. To address this challenge, a variety of approaches have been taken, ranging from instrumentation development to image post-processing. An example of the latter is deconvolution, where images are numerically deblurred based on a knowledge of the microscope point spread function. However, deconvolution can easily lead to noise-amplification artifacts. Deblurring by post-processing can also lead to negativities or fail to conserve local linearity between sample and image. We describe here a simple image deblurring algorithm based on pixel reassignment that inherently avoids such artifacts and can be applied to general microscope modalities and fluorophore types. Our algorithm helps distinguish nearby fluorophores even when these are separated by distances smaller than the conventional resolution limit, helping facilitate, for example, the application of single-molecule localization microscopy in dense samples. We demonstrate the versatility and performance of our algorithm under a variety of imaging conditions.

SUBMITTER: Zhao B

PROVIDER: S-EPMC10402078 | biostudies-literature | 2023 Jul

REPOSITORIES: biostudies-literature

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Resolution enhancement with deblurring by pixel reassignment (DPR).

Zhao Bingying B Mertz Jerome J

bioRxiv : the preprint server for biology 20230916

Improving the spatial resolution of a fluorescence microscope has been an ongoing challenge in the imaging community. To address this challenge, a variety of approaches have been taken, ranging from instrumentation development to image post-processing. An example of the latter is deconvolution, where images are numerically deblurred based on a knowledge of the microscope point spread function. However, deconvolution can easily lead to noise-amplification artifacts. Deblurring by post-processing ...[more]

PMID: 37546886

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Project description:PurposePrevious efforts at increasing spatial resolution have relied on decreasing focal spot and or detector element size. Many "super resolution" methods require physical movement of a component of the imaging system. This work describes a method for achieving spatial resolution on a scale smaller than the detector pixel without motion of the object or detector.MethodsWe introduce a weighting of the photon energy spectrum on a length scale smaller than a single pixel using a physical filter that can be placed between the focal spot and the object, between the object and the detector, or integrated into the x-ray source or detector. We refer to the method as sub pixel encoding (SPE). We show that if one acquires multiple measurements (i.e. x-ray projections), information can be synthesized at a spatial scale defined by the spectrum modulation, not the detector element size. Specifically, if one divides a detector pixel into n sub regions, and m photon-matter interactions are present, the number of x-ray measurements needed to solve for the detector response of each sub region is mxn. We discuss realizations of SPE using multiple x-ray spectra with an energy integrating detector, a single spectra with a photon counting detector, and the single photon-matter interaction case. We demonstrate the feasibility of the approach using a simulated energy integrating detector with a detector pitch of 2 mm for 80-140 kV medical and 200-600 kV industrial applications. Phantoms used for both example SPE realization had some features only a 1 mm detector could resolve. We calculate the covariance matrix of SPE output to characterize the and noise propagation and correlation of our test examples.ResultsThe mathematical foundation of SPE is provided, with details worked out for several detector types and energy ranges. Two numerical simulations were provided to demonstrate feasibility. In both the medical and industrial simulations, some phantom features were only observable with the 1 mm and SPE synthesized 2 mm detector, while the 2 mm detector was not able to visualize them. Covariance matrix analysis demonstrated negative diagonal terms for both example cases.ConclusionsThe concept of encoding object information at a length scale smaller than a single pixel element, and then retrieving that information was introduced. SPE simultaneously allows for an increase in spatial resolution and provides "dual energy" like information about the underlying photon-matter interactions.

Dataset Information

Resolution enhancement with deblurring by pixel reassignment (DPR).

Publications

Resolution enhancement with deblurring by pixel reassignment (DPR).

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