Hochschule Aalen, Institut fur Angewandte Forschung, Aalen, Germany.
J Biomed Opt. 2010 Jul-Aug;15(4):046017. doi: 10.1117/1.3470446.
Methods of wide-field fluorescence microscopy for measuring membrane dynamics of living cells are described, including spectral imaging as well as anisotropy imaging of the membrane marker 6-dodecanoyl-2-dimethylamino naphthalene (laurdan). Plasma membranes are selected by illumination with an evanescent electromagnetic field and distinguished from intracellular membranes assessed by whole-cell illumination. While fluorescence spectra of laurdan appeared red-shifted with decreasing membrane stiffness, fluorescence anisotropy and rotational correlation times were reduced with increasing membrane fluidity. Membrane stiffness was found to increase with decreasing temperature and increasing amounts of cholesterol and was always higher for the plasma membrane than for intracellular membranes. These effects may have some clinical relevance in the research of drug resistance or cell aging.
描述了用于测量活细胞膜动力学的宽场荧光显微镜方法，包括膜标记物 6-十二烷酰基-2-二甲基氨基萘（Laurdan）的光谱成像以及各向异性成像。通过渐逝电磁场所进行的照明选择质膜，并通过对整个细胞的照明来区分细胞内膜。随着膜硬度的降低，Laurdan 的荧光光谱出现红移，而荧光各向异性和旋转相关时间随着膜流动性的增加而降低。发现膜硬度随温度降低、胆固醇含量增加而增加，并且质膜的硬度总是高于细胞内膜。这些效应在研究药物抗性或细胞衰老时可能具有一定的临床意义。
