Tuning axial and lateral localization precision in 3D super-resolution microscopy with variable astigmatism.
Tuning axial and lateral localization precision in 3D super-resolution microscopy with variable astigmatism.
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DOI：
10.1364/ol.466213
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发表时间：
2022-11-01
期刊：
影响因子：
3.6
通讯作者：
中科院分区：
文献类型：
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散光成像是一种三维(3D)单分子荧光显微镜方法，它从一张图像中产生快速时间尺度上的超分辨率空间信息。它非常适合于分辨亚微米尺度的结构和毫秒范围内的时间行为。虽然传统的散光成像使用柱面透镜，但自适应光学使散光能够针对实验进行调整。我们在这里演示了、和中的精度是如何相互关联的，并随像散、位置和光子能级的变化而变化。这种实验驱动和验证的方法为生物成像策略中的散光选择提供了指导。
Astigmatism imaging is a three-dimensional (3D) single molecule fluorescence microscopy approach that yields super-resolved spatial information on a rapid timescale from a single image. It is ideally suited for resolving structures on a sub-micrometer scale and temporal behavior in the millisecond regime. While traditional astigmatism imaging utilizes a cylindrical lens, adaptive optics enables the astigmatism to be tuned for the experiment. We demonstrate here how the precisions in , , and are inter-linked and vary with the astigmatism, -position, and photon level. This experimentally-driven and -verified approach provides a guide for astigmatism selection in biological imaging strategies.