意大利技术研究所Vicidomini, Giuseppe团队研究了激光扫描显微镜超分辨光学切片的结构检测。2025年6月5日，《自然—光子学》杂志发表了这一成果。

快速探测器阵列能够有效地实现图像扫描显微镜，克服了共聚焦显微镜的空间分辨率和信噪比之间的权衡。然而，目前的图像扫描显微镜方法不提供光学切片，除非探测器尺寸有限，否则在厚样品上会失败，从而在光学切片和信噪比之间引入了一种新的权衡。 

研究组提出了一种克服这种限制的方法。通过单平面采集，重建了具有数字和光学超分辨率、高信噪比和增强光学切片的图像。基于对探测器阵列成像固有地嵌入轴向信息的观察，他们设计了一种直接的重建算法，该算法反转了图像扫描显微镜图像形成的物理模型。他们提出了一个全面的理论框架，并使用配备单光子雪崩二极管阵列探测器的定制装置捕获的生物样本图像验证了该方法。

研究组通过在线性和非线性区域激发荧光发射来证明该方法的可行性。此外，他们还推广了荧光寿命成像的算法，充分利用了单光子雪崩二极管阵列探测器的单光子计时能力。该方法优于传统的重建技术，可以扩展到任何激光扫描显微镜技术。

附：英文原文

Title: Structured detection for simultaneous super-resolution and optical sectioning in laser scanning microscopy

Author: Zunino, Alessandro, Garr, Giacomo, Perego, Eleonora, Zappone, Sabrina, Donato, Mattia, Vastenhouw, Nadine, Vicidomini, Giuseppe

Issue&Volume: 2025-06-05

Abstract: Fast detector arrays enable an effective implementation of image scanning microscopy, which overcomes the trade-off between spatial resolution and signal-to-noise ratio of confocal microscopy. However, current image scanning microscopy approaches do not provide optical sectioning and fail with thick samples unless the detector size is limited, thereby introducing a new trade-off between optical sectioning and signal-to-noise ratio. Here we propose a method that overcomes such a limitation. From single-plane acquisition, we reconstruct an image with digital and optical super-resolution, high signal-to-noise ratio and enhanced optical sectioning. On the basis of the observation that imaging with a detector array inherently embeds axial information, we designed a straightforward reconstruction algorithm that inverts the physical model of image scanning microscopy image formation. We present a comprehensive theoretical framework and validate our method with images of biological samples captured using a custom setup equipped with a single-photon avalanche diode array detector. We demonstrate the feasibility of our approach by exciting fluorescence emission in both linear and nonlinear regimes. Moreover, we generalize the algorithm for fluorescence lifetime imaging, fully exploiting the single-photon timing ability of the single-photon avalanche diode array detector. Our method outperforms conventional reconstruction techniques and can be extended to any laser scanning microscopy technique.

DOI: 10.1038/s41566-025-01695-0

Source: https://www.nature.com/articles/s41566-025-01695-0