Opt Lett. 2022 Feb 15;47(4):989-992. doi: 10.1364/OL.450973.
Two-photon microscopy (TPM) has provided critical in situ and in vivo information in biomedical studies due to its high resolution, intrinsic optical sectioning, and deep penetration. However, its relatively small field of view (FOV), which is usually determined by objectives, restricts its wide application. In this paper, we propose a segment-scanning sensorless adaptive optics method to extend the FOV and achieve high-resolution and large-FOV two-photon imaging. We demonstrated the proposed method by imaging fluorescent beads, cerebral nerve cells of mouse brain slices, and cerebral vasculature and microglia of live mice. The method extended the FOV of a commercial objective from 1.8 to 3.46 mm while maintaining a lateral resolution of 840 nm and high signal-to-noise ratio. Our technology is compatible with a standard TPM and can be used for large-scale biological exploration.
双光子显微镜（TPM）由于其高分辨率、固有光学切片和深穿透能力，为生物医学研究提供了关键的原位和在体信息。然而，其相对较小的视场（FOV），通常由物镜决定，限制了其广泛应用。在本文中，我们提出了一种分段扫描无传感器自适应光学方法来扩展 FOV 并实现高分辨率和大 FOV 双光子成像。我们通过对荧光珠、小鼠脑切片的脑神经细胞以及活体小鼠的脑血管和小胶质细胞进行成像来验证了该方法。该方法将商用物镜的 FOV 从 1.8 扩展到 3.46mm，同时保持 840nm 的横向分辨率和高信噪比。我们的技术与标准 TPM 兼容，可用于大规模的生物探索。
