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高对比光极化显微镜通过双标记的可光切换光蛋白进行高对比光极化

Lukas J Münker¹, Manuel Hohgardt¹, Andreas Albrecht¹

¹Technical University of Braunschweig, Braunschweig, Germany.

|July 3, 2025

概括

通过使用双重标记严格固光蛋白,可以在光极化显微镜 (FPM) 中增强对比度. 一种新的成像技术,FrExPAN,进一步提高了活细胞的定向选择性.

科学领域:

细胞成像细胞成像
显微镜技术的使用方法
生物物理学的生物物理.

背景情况:

光极化显微镜 (FPM) 受到光探针的旋转自由度的限制.
提高FPM的对比度需要更好地控制光体的方向.

研究的目的:

开发一种方法,使光蛋白 (FPs) 在活细胞中刚性固定.
提高FPM中的对比度和定向选择性.
引入一种用于增强偏振测量的新型成像方案.

主要方法:

利用可逆光切换光蛋白 (dt-rsFPs) 的双标记用于细胞结构.
开发了一种相机隔开关脉冲方案 (FrExPAN) 用于极化角度缩小.
应用FrExPAN成像到活的HeLa细胞和海马神经元的膜.

主要成果:

通过 dt-rsFPs 的刚性定,在 FPM 中显著增强了对比度.
成功实现了FrExPAN,有效地缩小了活细胞中的激发角度范围.
展示了具有明显方向的细胞结构的选择性成像.

结论:

通过双重标记对FP的刚性定是提高FPM对比度的广泛适用原则.
FrExPAN成像能够精确地选择细胞组件的方向.
该方法对定向对比成像和超分辨率技术 (如SPoD) 有潜在的影响.

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