Abstract
High-resolution and wide field-of-view (FOV) microscopic imaging plays a central role in diverse applications such as high-throughput screening and digital pathology. However, conventional microscopes face inherent trade-offs between the spatial resolution and FOV, which are fundamental limited by the space-bandwidth product (SBP) of the optical system. The resolution-FOV tradeoff can be effectively decoupled in Fourier ptychography microscopy (FPM), however, to date, the effective imaging NA achievable with a typical FPM system is still limited to the range of 0.4–0.7. Herein, we report, for the first time, a high-NA illumination based resolution-enhanced FPM (REFPM) platform, in which a LED-array-based digital oil-immersion condenser is used to create high-angle programmable plane-wave illuminations, endowing a 10×, 0.4 NA objective lens with final effective imaging performance of 1.6 NA. With REFPM, we present the highest-resolution results with a unprecedented half-pitch resolution of 154 nm at a wavelength of 435 nm across a wide FOV of 2.34 mm2, corresponding to an SBP of 98.5 megapixels (~50 times higher than that of the conventional incoherent microscope with the same resolution). Our work provides an important step of FPM towards high-resolution large-NA imaging applications, generating comparable resolution performance but significantly broadening the FOV of conventional oil-immersion microscopes.
Highlights
High-resolution wide-field imaging is essential for various applications in different fields, such as biological, biomedical research and digital pathology, which require large space-bandwidth product (SBP) to provide computational and statistical analyses for thousands of cells simultaneously across a wide field-of-view (FOV)[1,2,3,4]
We evaluate the performance of the resolution-enhanced FPM (REFPM)
We report a high-NA illumination based resolution-enhanced Fourier ptychography microscopy (FPM) platform, named REFPM, to generate high-resolution large-SBP reconstructions from a series of low-resolution raw images with the help of a 1.2 NA oil-immersion condenser and a 0.4 NA objective lens
Summary
High-resolution wide-field imaging is essential for various applications in different fields, such as biological, biomedical research and digital pathology, which require large space-bandwidth product (SBP) to provide computational and statistical analyses for thousands of cells simultaneously across a wide field-of-view (FOV)[1,2,3,4]. Illumination angles are scanned sequentially with a programmable light-emitting diode (LED) array, shifting different amounts of high spatial frequency information into the low NA objective lens This resolution improvement is analogous to coherent aperture synthesis[8, 9, 15,16,17,18] and structured-illumination[5, 6, 19,20,21,22] imaging. The reconstructed results indicate that REFPM achieves a large SBP nearly 50 times higher than that of the conventional incoherent microscope with the same resolution This unique technique could generate comparable resolution performance and significantly broaden the FOV of conventional oil-immersion microscopes, which would provide an important step of FPM towards high-resolution large-NA imaging applications
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