Figure 2—figure supplement 3. | Optical electrophysiology for probing function and pharmacology of voltage-gated ion channels

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Optical electrophysiology for probing function and pharmacology of voltage-gated ion channels

Figure 2—figure supplement 3.

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Harvard University, United States; Howard Hughes Medical Institute, Harvard University, United States
Figure 2—figure supplement 3.
Download figureOpen in new tabFigure 2—figure supplement 3. Cell-to-cell variability in Optopatch measurements.

(A) Nav1.7-OS HEK cells in a confluent monolayer were imaged in the presence of 3 μM amitriptyline with a 60× oil immersion objective (numerical aperture 1.45). Cells were illuminated with a red laser at 400 W/cm2. A blue prepulse (500 ms, 50 mW/cm2) depolarized membrane potential and allowed drug binding. A variable recovery period (40 ms to 5120 ms) led to partial unbinding. A blue test pulse (20 ms, 50 mW/cm2) probed the voltage spike induced by residual NaV1.7 capacity. Traces show fluorescence of QuasAr2 from five different single cells recorded in parallel. (B) Ratio of spike amplitude in test pulse to spike amplitude at the longest recovery time (5120 ms), as a function of recovery time for the five single cells shown in (A).

DOI: http://dx.doi.org/10.7554/eLife.15202.007