DOI: 10.1101/473843Nov 19, 2018Paper

Tethered Bichromophoric Fluorophore Quencher Voltage Sensitive Dyes

BioRxiv : the Preprint Server for Biology
Ping YanLeslie M Loew

Abstract

Voltage sensitive dyes (VSDs) are used for in vitro drug screening and for in vivo imaging of patterns of electrical activity. However, wide application of this technology is limited by poor sensitivity. A promising approach uses a 2-component system consisting of charged membrane permeable quenchers together with fluorophores labeling one side of the membrane; this produces voltage-dependent fluorescence quenching. However, to achieve good sensitivity, the quencher compound must be used at high concentrations, which can perturb the membrane capacitance or have other pharmacological effects. By developing tethered bichromophoric fluorophore quencher (TBFQ) dyes, where the fluorophore and quencher are covalently connected by a long hydrophobic chain, the concentration required is minimized, and the sensitivity is maximized. A series of 13 TBFQ dyes based on the Ami-noNaphthylEthenylPyridinium (ANEP) fluorophore and the dipicrylamine anion (DPA) quencher have been synthesized and tested in an artificial lipid bilayer apparatus. The best one from the screening, TBFQ1, shows a 2.5 fold change in fluorescence per 100mV change in membrane potential, and the response kinetics is in 10-20 ms range. This sensitivity is an order of magni...Continue Reading

Related Concepts

Diagnostic Imaging
Dyes
Fluorescent Dyes
Ions
Lipid Bilayers
Molecular Probe Techniques
Dipicrylamine
Drug Screen (Procedure)
Membrane
Fluorophore

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