Random-access wide-field mesoscopy for centimetre-scale imaging of biodynamics with subcellular resolution
Ruheng Shi, Xinyue Chen, Junhao Deng, Junhao Liang, Kuikui Fan, Feifan Zhou, Peifu Tang, Licheng Zhang, Lingjie Kong
Abstract
Abstract Benefitting from the advantages of high imaging throughput and low cost, wide-field microscopy has become indispensable in biomedical studies. However, it remains challenging to record biodynamics with a large field of view and high spatiotemporal resolution due to the limited space–bandwidth product. Here we propose random-access wide-field (RA-WiFi) mesoscopy for the imaging of in vivo biodynamics over a 163.84 mm 2 area with a spatial resolution of ~2.18 μm. We extend the field of view beyond the nominal value of the objective by enlarging the object distance, which leads to a lower field angle, followed by the correction of optical aberrations. We also implement random-access scanning with structured illumination, which enables optical-sectioning capability and high imaging contrast. The multi-plane imaging capability also makes the technique suitable for curved-surface samples. We demonstrate RA-WiFi mesoscopy in multi-modal imaging, including bright-field, dark-field and multi-colour fluorescence imaging. Specifically, we apply RA-WiFi mesoscopy to calcium imaging of cortex-wide neural network activities in awake mice in vivo, under both physiological and pathological conditions. We also show its unique capability in the three-dimensional random access of irregular regions of interest via the biodynamic imaging of mouse spinal cords in vivo. As a compact, low-cost mesoscope with optical-sectioning capability, RA-WiFi mesoscopy will enable broad applications in the biodynamic study of biological systems.