What question did this study set out to answer?

This research aims to enhance volumetric imaging techniques using a nonparaxial self-bending beam for improved depth-of-field.

June 19, 2026

Extended depth-of-field volumetric microscopy based on bidirectionally separated nonparaxial self-bending beam

Key Points

This research aims to enhance volumetric imaging techniques using a nonparaxial self-bending beam for improved depth-of-field.
Engineered a nonparaxial self-bending beam with depth encoding using cubic phase and asymmetric chirp.
Conducted experiments on standard samples and biological tissues to validate imaging fidelity and accuracy.
Implemented fluorescence microscopy with high numerical aperture to achieve extended depth of field.
Achieved an extended depth-of-field of 70 μm in a 100×, NA 1.45 fluorescence microscope.
Demonstrated near-diffraction-limited resolution and 3D imaging fidelity over approximately 40 μm depth.
Validated applicability through imaging of a 16-μm-thick mouse kidney section.

Abstract

We propose a detection-side volumetric imaging strategy based on a nonparaxial self-bending beam (SBB), achieving an extended depth-of-field (DOF) of 70 μm in a 100×, numerical-aperture (NA) 1.45 fluorescence microscope. By combining cubic phase, asymmetric chirp, and nonparaxial residual-phase compensation, we engineer a nonparaxial SBB with robust depth encoding under high-NA focusing. With bidirectional separation, the nonparaxial SBB enables high-resolution 3D imaging. A standard-sample experiment validates the 3D imaging fidelity and reconstruction accuracy over a DOF of ∼40 μm. A volumetric fluorescent beads experiment demonstrates the ∼70 μm DOF and the near-diffraction-limited resolution. Imaging of a 16-μm-thick mouse kidney section further evaluates the applicability of the method to continuous biological tissue. This framework provides a practical paradigm for deploying structured beams to enhance imaging performance in high-NA volumetric microscopes.

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