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基于双螺旋点扩散函数工程的散射介质中的成像和定位。

Imaging and positioning through scattering media with double-helix point spread function engineering.

机构信息

Chinese Academy of Sciences, Shanghai Institute of Optics and Fine Mechanics, Key Laboratory of Quantum Optics, Shanghai, China.

University of Chinese Academy of Sciences, Center of Materials Science and Optoelectronics Engineering, Beijing, China.

出版信息

J Biomed Opt. 2023 Apr;28(4):046008. doi: 10.1117/1.JBO.28.4.046008. Epub 2023 Apr 25.

DOI:10.1117/1.JBO.28.4.046008
PMID:37114201
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10127513/
Abstract

SIGNIFICANCE

Double-helix point spread function (DH-PSF) microscopy has been developed for three-dimensional (3D) localization and imaging at super-resolution but usually in environments with no or weak scattering. To date, super-resolution imaging through turbid media has not been reported.

AIM

We aim to explore the potential of DH-PSF microscopy in the imaging and localization of targets in scattering environments for improved 3D localization accuracy and imaging quality.

APPROACH

The conventional DH-PSF method was modified to accommodate the scanning strategy combined with a deconvolution algorithm. The localization of a fluorescent microsphere is determined by the center of the corresponding double spot, and the image is reconstructed from the scanned data by deconvoluting the DH-PSF.

RESULTS

The resolution, i.e., the localization accuracy, was calibrated to 13 nm in the transverse plane and 51 nm in the axial direction. Penetration thickness could reach an optical thickness (OT) of 5. Proof-of-concept imaging and the 3D localization of fluorescent microspheres through an eggshell membrane and an inner epidermal membrane of an onion are presented to demonstrate the super-resolution and optical sectioning capabilities.

CONCLUSIONS

Modified DH-PSF microscopy can image and localize targets buried in scattering media using super-resolution. Combining fluorescent dyes, nanoparticles, and quantum dots, among other fluorescent probes, the proposed method may provide a simple solution for visualizing deeper and clearer in/through scattering media, making super-resolution microscopy possible for various demanding applications.

摘要

意义

双螺旋点扩散函数 (DH-PSF) 显微镜已被开发用于在超分辨率下进行三维 (3D) 定位和成像，但通常在无散射或弱散射的环境中。迄今为止，尚未有通过混浊介质进行超分辨率成像的报道。

目的

我们旨在探索 DH-PSF 显微镜在散射环境中对目标进行成像和定位的潜力，以提高 3D 定位精度和成像质量。

方法

对传统的 DH-PSF 方法进行了修改，以适应与去卷积算法相结合的扫描策略。通过对双点对应的中心点进行定位，确定荧光微球的位置，然后通过对 DH-PSF 进行去卷积，从扫描数据中重建图像。

结果

分辨率（即定位精度）在横向平面上校准为 13nm，在轴向方向上校准为 51nm。穿透厚度可达到 5 个光学厚度 (OT)。通过鸡蛋壳膜和洋葱内表皮膜对荧光微球进行了超分辨率成像和 3D 定位的概念验证，证明了该显微镜具有超分辨率和光学切片的能力。

结论

经改进的 DH-PSF 显微镜可在散射介质中对目标进行超分辨率成像和定位。通过结合荧光染料、纳米粒子和量子点等荧光探针，该方法可为可视化更深、更清晰的散射介质提供简单的解决方案，使超分辨率显微镜能够应用于各种具有挑战性的应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8331/10127513/2f2f6077f280/JBO-028-046008-g010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8331/10127513/9359a880df47/JBO-028-046008-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8331/10127513/945d9adb8c20/JBO-028-046008-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8331/10127513/c0474c19706e/JBO-028-046008-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8331/10127513/885266148a8d/JBO-028-046008-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8331/10127513/79da2e0b81fc/JBO-028-046008-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8331/10127513/e5ae163872a7/JBO-028-046008-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8331/10127513/c5eca09a59e9/JBO-028-046008-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8331/10127513/c54b0e93d55d/JBO-028-046008-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8331/10127513/28f63c229d28/JBO-028-046008-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8331/10127513/2f2f6077f280/JBO-028-046008-g010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8331/10127513/9359a880df47/JBO-028-046008-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8331/10127513/945d9adb8c20/JBO-028-046008-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8331/10127513/c0474c19706e/JBO-028-046008-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8331/10127513/885266148a8d/JBO-028-046008-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8331/10127513/79da2e0b81fc/JBO-028-046008-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8331/10127513/e5ae163872a7/JBO-028-046008-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8331/10127513/c5eca09a59e9/JBO-028-046008-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8331/10127513/c54b0e93d55d/JBO-028-046008-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8331/10127513/28f63c229d28/JBO-028-046008-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8331/10127513/2f2f6077f280/JBO-028-046008-g010.jpg

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