Configurable fiber-bundle–relay scanning oblique plane microscopy for in vivo volumetric imaging

Light sheet fluorescence microscopy, including oblique plane microscopy (OPM), enables high-speed volumetric imaging but is often constrained by complex and rigid image-relay optics that limit flexibility in system configuration and sample access. Here, we present a configurable scanning OPM that employs a coherent fiber-bundle image relay to decouple a compact front-end imaging unit from a stationary detection module. An angle-polished, magnified coupling scheme, together with a fiber-pattern removal algorithm, mitigates core-induced pixelation while preserving volumetric imaging performance. The system achieves volumetric imaging over a 300×300×50µm 3 field of view at 2volumes⋅s −1 maximum, with effective lateral resolution on the order of 1 µm and axial resolution on the order of 2 µm after full three-dimensional reconstruction. Performance is validated through ex vivo imaging of mouse cornea, brain, and colon, as well as in vivo longitudinal imaging in corneal injury models. Time-lapse 3D tracking of inflammatory cells within the corneal stroma further demonstrates the system’s capability for dynamic cellular imaging. By combining intrinsic optical sectioning with a compact and configurable architecture compatible with clinically relevant imaging scenarios, this approach extends the applicability of scanning OPM toward translational biomedical imaging.