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 mm² 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.

受益于高成像通量和低成本的优势，宽视场显微镜已成为生物医学研究中不可或缺的一部分。然而，由于空间带宽积有限，以大视场和高时空分辨率记录生物动力学仍然具有挑战性。在这里，我们提出了随机访问宽视场（RA-WiFi）介观镜，用于在 163.84 mm ²区域内进行体内生物动力学成像，空间分辨率约为 2.18 μm。我们通过增大物距来将视场扩展到物镜标称值之外，从而降低视场角，然后进行光学像差校正。我们还利用结构照明实现随机访问扫描，从而实现光学切片功能和高成像对比度。多平面成像能力也使得该技术适用于曲面样品。我们展示了 RA-WiFi 介观镜在多模态成像中的应用，包括明场、暗场和多色荧光成像。具体来说，我们将 RA-WiFi 介观镜应用于生理和病理条件下清醒小鼠体内皮质范围神经网络活动的钙成像。我们还通过体内小鼠脊髓的生物动力学成像展示了其在三维随机访问不规则感兴趣区域方面的独特能力。作为一款具有光学切片功能的紧凑、低成本介观镜，RA-WiFi 介观镜将在生物系统的生物动力学研究中实现广泛的应用。