Kavli Affiliate: Changhuei Yang
| First 5 Authors: Haowen Zhou, Haowen Zhou, , ,
| Summary:
Automation in optical microscopy is critical for enabling high-throughput
imaging across a wide range of biomedical applications. Among the essential
components of automated systems, robust autofocusing plays a pivotal role in
maintaining image quality for both single-plane and volumetric imaging.
However, conventional autofocusing methods often struggle with implementation
complexity, limited generalizability across sample types, incompatibility with
thick specimens, and slow feedback. We recently discovered a phenomenon that
the digitally summed Fourier spectrum of two images acquired from two-angle
illumination exhibits interference-like fringe modulation when the sample is
defocused. These digital fringes correlate directly with defocus through a
physics-based relation. Based on this principle, we developed an automatic,
efficient, and generalizable defocus detection method termed digital defocus
aberration interference (DAbI). Implemented with a simple two-LED setup, DAbI
can quantify the defocus distance over a range of 212 times the depth-of-field
(DoF) for thin samples and 300 times for thick specimens. It can additionally
extend the natural DoF of the imaging system by 20 folds when integrated with
complex-field imaging. We demonstrated the versatile applications of DAbI on
brightfield, complex-field, refractive index, confocal, and widefield
fluorescence imaging, establishing it as a promising solution for automated,
high-throughput optical microscopy.
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