【2h】

Palette of fluorinated voltage-sensitive hemicyanine dyes

机译:氟化电压敏半菁染料的调色板

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摘要

Optical recording of membrane potential permits spatially resolved measurement of electrical activity in subcellular regions of single cells, which would be inaccessible to electrodes, and imaging of spatiotemporal patterns of action potential propagation in excitable tissues, such as the brain or heart. However, the available voltage-sensitive dyes (VSDs) are not always spectrally compatible with newly available optical technologies for sensing or manipulating the physiological state of a system. Here, we describe a series of 19 fluorinated VSDs based on the hemicyanine class of chromophores. Strategic placement of the fluorine atoms on the chromophores can result in either blue or red shifts in the absorbance and emission spectra. The range of one-photon excitation wavelengths afforded by these new VSDs spans 440–670 nm; the two-photon excitation range is 900–1,340 nm. The emission of each VSD is shifted by at least 100 nm to the red of its one-photon excitation spectrum. The set of VSDs, thus, affords an extended toolkit for optical recording to match a broad range of experimental requirements. We show the sensitivity to voltage and the photostability of the new VSDs in a series of experimental preparations ranging in scale from single dendritic spines to whole heart. Among the advances shown in these applications are simultaneous recording of voltage and calcium in single dendritic spines and optical electrophysiology recordings using two-photon excitation above 1,100 nm.
机译:膜电位的光学记录可以对单个细胞的亚细胞区域的电活动进行空间分辨测量,而这是电极无法到达的,并且可以对动作电位在时空模式中在可兴奋性组织(例如大脑或心脏)中的传播进行成像。但是,可用的压敏染料(VSD)并不总是与用于感测或操纵系统生理状态的新可用光学技术在光谱上兼容。在这里,我们根据生色团的半花菁类描述了一系列的19个氟化VSD。氟原子在发色团上的战略性放置可能导致吸收光谱和发射光谱发生蓝移或红移。这些新的VSD提供的单光子激发波长范围跨越440-670 nm。双光子激发范围为900–1,340 nm。每个VSD的发射至少移动100 nm到其单光子激发光谱的红色。因此,这套VSD为光学记录提供了扩展的工具包,可满足广泛的实验要求。我们在一系列实验准备中显示了对新VSD的电压敏感性和光稳定性,这些实验准备范围从单个树突棘到整个心脏。在这些应用中显示的进步包括在单个树突棘中同时记录电压和钙,以及使用1100 nm以上的双光子激发进行光学电生理记录。

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