近日，清华大学的孔令杰及其研究小组与中国人民解放军总医院的张里程&唐佩福等人合作并取得一项新进展。经过不懈努力，他们提出用于具有亚细胞分辨率生物动力学厘米级成像的随机存取宽视场介观显微镜。相关研究成果已于2024年4月17日在国际知名学术期刊《自然—光子学》上发表。
该研究团队提出了随机存取宽视场(RA-WiFi)介观显微镜，旨在实现163.84mm²区域内的活体生物动力学成像，其空间分辨率高达约2.18μm。研究人员通过扩大物镜的距离，使视场超出物镜的标称值，从而降低视场角度，然后进行光学像差的校正。研究人员还实现了随机访问扫描与结构化照明技术，显著提升了光学切片能力和成像对比度。多平面成像能力也使该技术适用于曲面样品。他们展示了RA-WiFi介观显微镜在多模态成像中的应用，包括明场、暗场和多色荧光成像。
具体而言，他们应用RA-WiFi介观显微镜对活体中清醒小鼠生理和病理条件下的，全皮质神经网络活动进行钙成像。此外，他们还通过对小鼠脊髓在三维随机访问的不规则感兴趣区域内进行生物动力学成像，进一步证明了该技术的独特优势。作为一种具有光学切片能力的紧凑、低成本的介观显微镜，RA-WiFi介显微观镜将在生物系统的生物动力学研究中得到广泛应用。
据悉，宽视场显微镜具有成像通量高、成本低等优点，已成为生物医学研究中不可或缺的技术。然而，由于有限的空间带宽乘积，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。
附：英文原文
Title: Random-access wide-field mesoscopy for centimetre-scale imaging of biodynamics with subcellular resolution
Author: Shi, Ruheng, Chen, Xinyue, Deng, Junhao, Liang, Junhao, Fan, Kuikui, Zhou, Feifan, Tang, Peifu, Zhang, Licheng, Kong, Lingjie
Issue&Volume: 2024-04-17
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 163.84mm2 area with a spatial resolution of ~2.18μm. 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.
DOI: 10.1038/s41566-024-01422-1
Source: https://www.nature.com/articles/s41566-024-01422-1