Department of Chemistry and Biochemistry, University of California, Los Angeles, California 90095-1569, United States.
Department of Mechanical Engineering, University of California, Santa Barbara, California 93106-5200, United States.
J Am Chem Soc. 2020 Sep 16;142(37):16072-16081. doi: 10.1021/jacs.0c07761. Epub 2020 Sep 4.
The bioorthogonal nature of perfluorocarbons provides a unique platform for introducing dynamic nano- and microdroplets into cells and organisms. To monitor the localization and deformation of the droplets, fluorous soluble fluorophores that are compatible with standard fluorescent protein markers and applicable to cells, tissues, and small organisms are necessary. Here, we introduce fluorous cyanine dyes that represent the most red-shifted fluorous soluble fluorophores to date. We study the effect of covalently appended fluorous tags on the cyanine scaffold and evaluate the changes in photophysical properties imparted by the fluorous phase. Ultimately, we showcase the utility of the fluorous soluble pentamethine cyanine dye for tracking the localization of perfluorocarbon nanoemulsions in macrophage cells and for measurements of mechanical forces in multicellular spheroids and zebrafish embryonic tissues. These studies demonstrate that the red-shifted cyanine dyes offer spectral flexibility in multiplexed imaging experiments and enhanced precision in force measurements.
全氟碳化合物的生物正交性质为向细胞和生物体中引入动态纳米和微滴提供了独特的平台。为了监测液滴的定位和变形,需要使用与标准荧光蛋白标记物兼容且适用于细胞、组织和小型生物的可溶解于氟相的荧光团。在这里,我们引入了氟花青染料,它们代表了迄今为止红移程度最大的可溶解于氟相的荧光团。我们研究了共价连接的氟标签对花青支架的影响,并评估了氟相赋予的光物理性质的变化。最终,我们展示了可溶解于氟相的五甲川菁染料在追踪全氟碳纳米乳液在巨噬细胞中的定位以及测量多细胞球体和斑马鱼胚胎组织中的机械力方面的应用。这些研究表明,红移花青染料在多路复用成像实验中提供了光谱灵活性,并在力测量中提高了精度。