In situ correction of liquid meniscus in cell culture imaging system based on parallel Fourier ptychographic microscopy (96 Eyes)

Abstract

We collaborated with Amgen and spent five years in designing and fabricating next generation multi-well plate imagers based on Fourier ptychographic microscopy (FPM). A 6-well imager (Emsight) and a low-cost parallel microscopic system (96 Eyes) based on parallel FPM were reported in our previous work. However, the effect of liquid meniscus on the image quality is much stronger than anticipated, introducing obvious wavevector misalignment and additional image aberration. To this end, an adaptive wavevector correction (AWC-FPM) algorithm and a pupil recovery improvement strategy are presented to solve these challenges in situ. In addition, dual-channel fluorescence excitation is added to obtain structural information for microbiologists. Experiments are demonstrated to verify their performances. The accuracy of angular resolution with our algorithm is within 0.003 rad. Our algorithms would make the FPM algorithm more robust and practical and can be extended to other FPM-based applications to overcome similar challenges.

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