Complex structures in living cells and tissues induce wavefront errors when light waves pass through them, and images observed with optical microscopes are undesirably blurred. This problem is especially serious for living plant cells because images are strikingly degraded even within a single cell. Adaptive optics (AO) is expected to be a solution to this problem by correcting such wavefront errors, thus enabling high-resolution imaging. In particular, scene-based AO involves wavefront sensing based on the image correlation between subapertures in a Shack–Hartmann wavefront sensor and thus does not require an intense point light source. However, the complex 3D structures of living cells often cause low correlation between subimages, leading to loss of accuracy in wavefront sensing. This paper proposes a novel method for scene-based sensing using only image correlations between adjacent subapertures. The method can minimize changes between subimages to be correlated and thus prevent inaccuracy in phase estimation. Using an artificial test target mimicking the optical properties of a layer of living plant cells, an imaging performance with a Strehl ratio of approximately 0.5 was confirmed. Upon observation of chloroplast autofluorescence inside living leaf cells of the moss Physcomitrium patens, recovered resolution images were successfully obtained even with complex biological structures. Under bright-field illumination, the proposed method outperformed the conventional method, demonstrating the future potential of this method for label- and damage-free AO microscopy. Several points for improvement in terms of the effect of AO correction are discussed.

中文翻译：

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具有基于邻域相关性的波前传感的自适应光学显微镜

当光波穿过活细胞和组织中的复杂结构时，它们会产生波前误差，并且用光学显微镜观察到的图像会变得模糊。这个问题对于活的植物细胞来说尤其严重，因为即使在单个细胞内图像也会显着退化。自适应光学（AO）有望通过纠正此类波前误差来解决这一问题，从而实现高分辨率成像。特别是，基于场景的 AO 涉及基于 Shack-Hartmann 波前传感器中子孔径之间的图像相关性的波前传感，因此不需要强点光源。然而，活细胞复杂的 3D 结构通常会导致子图像之间的相关性较低，从而导致波前传感精度下降。本文提出了一种仅使用相邻子孔径之间的图像相关性的基于场景的传感的新方法。该方法可以最小化要相关的子图像之间的变化，从而防止相位估计的不准确。使用模拟活植物细胞层光学特性的人工测试目标，证实了斯特列尔比约为 0.5 的成像性能。通过观察小立碗藓活叶细胞内的叶绿体自发荧光，即使具有复杂的生物结构，也能成功获得恢复分辨率的图像。在明场照明下，所提出的方法优于传统方法，展示了该方法在无标记和无损伤 AO 显微镜方面的未来潜力。讨论了 AO 校正效果方面需要改进的几点。