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Abstract
Neurons communicate by using electrical signals, mediated by transient changes in the voltage across the plasma membrane. Optical techniques for visualizing these transmembrane potentials could revolutionize the field of neurobiology by allowing the spatial profile of electrical activity to be imaged in real time with high resolution, along individual neurons or groups of neurons within their native networks.1, 2 Second harmonic generation (SHG) is one of the most promising methods for imaging membrane potential, although so far this technique has only been demonstrated with a narrow range of dyes.3 Here we show that SHG from a porphyrin-based membrane probe gives a fast electro-optic response to an electric field which is about 5–10 times greater than that of conventional styryl dyes. Our results indicate that porphyrin dyes are promising probes for imaging membrane potential.
Highlights
The sensitivity of the porphyrin dye JR1 and the styryl dyes FM4-64, di-4-ANEPPS, and RH237 to transmembrane potential was investigated using the setup shown in Figure 1.[15,26,27] Each dye was added to an aqueous solution of phosphate buffered saline to give a dye concentration of 5 mm
We chose to compare JR1 directly with three previously studied styryl dyes, under identical conditions, because earlier work has shown that a dyes response to an electric field can depend on experimental parameters such as the composition of the lipid bilayer, the laser wavelength, and the time scale of the measurement.[3,6,19,30]
Our results for FM4-64, di-4ANEPPS, and RH237 are broadly in line with previously published studies on these dyes, this is the first time that all three dyes have been compared under identical conditions in HLBs
Summary
[c] R"v;s is the normalized crosscorrelation, which measures how well the dye reports the applied potential waveform (see Figure S2 in the Supporting Information).
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