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Updated: Jun 28, 2025

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Published on: December 9, 2013

时间聚焦多光子激发单分子定位显微镜使用自发闪的光灯

Jian-Zong Lai¹, Chun-Yu Lin², Shean-Jen Chen²

¹Department of Optics and Photonics, National Central University, No. 300, Zhongda Rd., Zhongli Dist., Taoyuan City, 32001, Taiwan.

Angewandte Chemie (International ed. in English)

|April 20, 2024

概括

这项研究引入了一种新的单分子局部化显微镜技术,使用时间聚焦多光子激发 (TFMPE) 进行高分辨率3D成像. 该方法提高了光子效率,并可视化亚细胞结构和阿尔茨海默氏症.

科学领域:

生物物理学的生物物理.
显微镜的使用方法
细胞生物学细胞生物学

背景情况:

单分子定位显微镜 (SMLM) 能够实现纳米尺度成像,但在厚的样本中面临着光漂白和光子预算的挑战.
多光子激发 (MPE) 为深层组织成像提供了优势,但需要对SMLM进行优化.

研究的目的:

开发和验证单波长时间聚焦多光子激发 (TFMPE) SMLM技术,以改进3D成像.
为了优化TPE波长,以提高从闪的光灯中检测光子的效果.
在厚厚的生物样本中可视化亚细胞结构和病理特征.

主要方法:

实现单波长TFMPE用于闪的光体的广场和轴局限的两光子激发 (TPE).
进行TPE光谱测量以确定最佳激发波长.
结合TFMPE-SMLM与形成像用于3D重建.

主要成果:

实现了纳米级空间分辨率 (约. 51nm) 和高定位精度 (18±6nm) 在癌细胞微管的二维SMLM中.
在阿尔茨海默病小鼠大脑组织中展示了粉样β沉积物的3DTFMPE-SMLM成像.
成功地减少了焦点外地区的光漂白效应,改善了光子预算的利用.

结论:

TFMPE-SMLM是一个强大的工具,用于高分辨率的3D成像子细胞结构和疾病病理在厚的标本.
优化的TPE波长显著提高了SMLM的光子检测效率.
这种技术为研究纳米级复杂生物系统提供了一个有前途的方法.

关键词:

粉样蛋白-β 沉积物单分子局部化显微镜.自发地闪的光.时间聚焦的多光子激发刺激.三维成像三维成像技术

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