We demonstrate a deep learning-based offline autofocusing method, termed Deep-R, to rapidly and blindly autofocus a single-shot microscopy image captured at an arbitrary out-of-focus plane. Deep-R is experimentally validated using various tissue sections that were imaged with fluorescence and brightfield microscopes. Furthermore, snapshot autofocusing under different defocusing scenarios is demonstrated, including uniform axial-defocusing, sample tilting, cylindrical and spherical distortions within the field-of-view. Compared with other online autofocusing algorithms, Deep-R is significantly faster while having comparable image performance. Deep-R framework will enable high-throughput microscopic imaging over large fields-of-view, improving the overall imaging throughput, also reducing the photon dose on the sample.
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