VoltageFluor 染料和荧光寿命成像技术用于膜电位的光学测量。
VoltageFluor dyes and fluorescence lifetime imaging for optical measurement of membrane potential.
机构信息
Department of Chemistry, University of California, Berkeley, CA, United States.
Department of Chemistry, University of California, Berkeley, CA, United States; Department of Molecular & Cell Biology, University of California, Berkeley, CA, United States; Helen Wills Neuroscience Institute, University of California, Berkeley, CA, United States.
出版信息
Methods Enzymol. 2021;653:267-293. doi: 10.1016/bs.mie.2021.02.009. Epub 2021 Mar 29.
Membrane potential is a fundamental biophysical parameter common to all of cellular life. Traditional methods to measure membrane potential rely on electrodes, which are invasive and low-throughput. Optical methods to measure membrane potential are attractive because they have the potential to be less invasive and higher throughput than classic electrode based techniques. However, most optical measurements rely on changes in fluorescence intensity to detect changes in membrane potential. In this chapter, we discuss the use of fluorescence lifetime imaging microscopy (FLIM) and voltage-sensitive fluorophores (VoltageFluors, or VF dyes) to estimate the millivolt value of membrane potentials in living cells. We discuss theory, application, protocols, and shortcomings of this approach.
膜电位是所有细胞生命共有的基本生物物理参数。传统的测量膜电位的方法依赖于电极，而这些电极具有侵入性且通量低。测量膜电位的光学方法很有吸引力，因为它们有可能比基于经典电极的技术具有更低的侵入性和更高的通量。然而，大多数光学测量依赖于荧光强度的变化来检测膜电位的变化。在本章中，我们将讨论使用荧光寿命成像显微镜 (FLIM) 和电压敏感荧光染料 (VoltageFluors，或 VF 染料) 来估计活细胞中膜电位的毫伏值。我们讨论了这种方法的理论、应用、方案和缺点。
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