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Published February 1, 2016 | Published
Journal Article Open

Wide field-of-view fluorescence image deconvolution with aberration-estimation from Fourier ptychography

Abstract

This paper presents a method to simultaneously acquire an aberration-corrected, wide field-of-view fluorescence image and a high-resolution coherent bright-field image using a computational microscopy method. First, the procedure applies Fourier ptychographic microscopy (FPM) to retrieve the amplitude and phase of a sample, at a resolution that significantly exceeds the cutoff spatial frequency of the microscope objective lens. At the same time, redundancy within the set of acquired FPM bright-field images offers a means to estimate microscope aberrations. Second, the procedure acquires an aberrated fluorescence image, and computationally improves its resolution through deconvolution with the estimated aberration map. An experimental demonstration successfully improves the bright-field resolution of fixed, stained and fluorescently tagged HeLa cells by a factor of 4.9, and reduces the error caused by aberrations in a fluorescence image by up to 31%, over a field of view of 6.2 mm by 9.3 mm. For optimal deconvolution, we show the fluorescence image needs to have a signal-to-noise ratio of at least ~18.

Additional Information

© 2016 Optical Society of America. Received 2 Nov 2015; revised 16 Dec 2015; accepted 29 Dec 2015; published 7 Jan 2016. This project was funded by the National Institute of Health (NIH) Agency Award: R01 AI096226; and the Caltech Innovation Initiative (CII): 25570015. We thank Mooseok Jang for helpful discussions.

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