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在光学显微镜中通过可信度珠的反射干涉度通过光学显微镜的纳米分辨率实现数小时的漂移校正

Binh Phan¹, Michael R Stoneman¹, Sabita Sharma¹

¹Department of Physics, University of Wisconsin-Milwaukee, 3135 N. Maryland Ave., Milwaukee, Wisconsin 53211, United States.

|February 4, 2025

概括

为增强垂直分辨率 (FREVR) 重新调整焦点使用信托珠精确跟踪焦点,使稳定的显微镜能够持续数小时. 这种技术显著提高了生物样本的长期成像.

科学领域:

生物物理学的生物物理.
光学显微镜的使用方法
细胞生物学细胞生物学

背景情况:

光学显微镜对于实时生物动态至关重要.
焦点漂移将显微镜持续时间限制在几分钟内,没有减轻.
长期成像需要精确的焦点稳定.

研究的目的:

引入用于增强垂直分辨率 (FREVR) 的焦点调整.
开发一种方法来克服显微镜中的焦点偏移.
提高长期显微镜中的时间和空间分辨率.

主要方法:

与标本一起使用微米大小的信托珠.
从珠子中追踪干扰边缘,用于纳米精度的焦点检测.
使用高速的CMOS和敏感的EMCCD摄像头进行珠子跟踪和光量化.

主要成果:

在几个小时内实现了稳定,聚焦的测量.
在单分子系统和哺乳动物细胞上证明有效.
成功成像标记的活性纤维和膜蛋白.

结论:

FREVR显著延长了显微镜的稳定性和持续时间.
该方法提供纳米级焦点精度.
FREVR具有广泛的潜力,可以推进显微镜技术.

关键词:

焦点调整调整的焦点调整.几个小时的活细胞动态.显微镜漂移估计微镜漂移估计光学纳米显微镜的使用方法

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