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使用动态样条 PSF 模型实现 4Pi-STORM 的最佳精度和准确性。

Optimal precision and accuracy in 4Pi-STORM using dynamic spline PSF models.

机构信息

Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.

Department of Optical Nanoscopy, Institute for NanoPhotonics, Göttingen, Germany.

出版信息

Nat Methods. 2022 May;19(5):603-612. doi: 10.1038/s41592-022-01465-8. Epub 2022 May 16.

DOI:10.1038/s41592-022-01465-8
PMID:35577958
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9119851/
Abstract

Coherent fluorescence imaging with two objective lenses (4Pi detection) enables single-molecule localization microscopy with sub-10 nm spatial resolution in three dimensions. Despite its outstanding sensitivity, wider application of this technique has been hindered by complex instrumentation and the challenging nature of the data analysis. Here we report the development of a 4Pi-STORM microscope, which obtains optimal resolution and accuracy by modeling the 4Pi point spread function (PSF) dynamically while also using a simpler optical design. Dynamic spline PSF models incorporate fluctuations in the modulation phase of the experimentally determined PSF, capturing the temporal evolution of the optical system. Our method reaches the theoretical limits for precision and minimizes phase-wrapping artifacts by making full use of the information content of the data. 4Pi-STORM achieves a near-isotropic three-dimensional localization precision of 2-3 nm, and we demonstrate its capabilities by investigating protein and nucleic acid organization in primary neurons and mammalian mitochondria.

摘要

使用两个物镜（4Pi 检测）进行相干荧光成像是一种能够在三维空间中实现亚 10nm 空间分辨率的单分子定位显微镜技术。尽管具有出色的灵敏度，但由于仪器复杂以及数据分析的挑战性，该技术的更广泛应用受到了阻碍。在这里，我们报告了一种 4Pi-STORM 显微镜的开发，该显微镜通过在动态建模 4Pi 点扩散函数 (PSF) 的同时使用更简单的光学设计，获得了最佳的分辨率和准确性。动态样条 PSF 模型纳入了实验确定的 PSF 调制相位的波动，从而捕获了光学系统的时间演化。我们的方法通过充分利用数据的信息含量，达到了精度的理论极限，并最大限度地减少了相位缠绕伪影。4Pi-STORM 实现了接近各向同性的三维定位精度，精度为 2-3nm，我们通过研究原代神经元和哺乳动物线粒体中的蛋白质和核酸组织来证明其性能。

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3
Accurate 4Pi single-molecule localization using an experimental PSF model.使用实验性点扩散函数模型进行精确的4Pi单分子定位
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4
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5
Super-Resolution Goes Viral: T4 Virus Particles as Versatile 3D-Bio-NanoRulers.超分辨率迅速传播：T4病毒颗粒作为多功能3D生物纳米尺
Adv Mater. 2025 Mar;37(12):e2403365. doi: 10.1002/adma.202403365. Epub 2025 Jan 16.
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4
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