Figure 2—figure supplement 4. | 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 4.

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Harvard University, United States; Howard Hughes Medical Institute, Harvard University, United States
Figure 2—figure supplement 4.
Download figureOpen in new tabFigure 2—figure supplement 4. Effects of intense red laser illumination.

(A) Heating by red laser illumination (635 nm, 400 W/cm2). Temperatures were recorded with a small thermocouple not directly illuminated by the laser. Measurements were performed in a single well of a 384-well plate with 36 μL buffer, and in a 35 mm dish with 2 mL buffer (error bars represent s.e.m., n = 4 replicates). (B) Test for photobleaching and phototoxicity. Nav1.7-OS HEK cell were illuminated with red laser (635 nm, 400 W/cm2) continuously for 10 min. Every one minute, the cells were stimulated with blue light (6 pulses of 20 ms, 5 Hz, 50 mW/cm2). The first, sixth and the tenth spike trains are shown above. (C) SNR and spike height (ΔF/F0) for each spike train in (B) as a function of red laser illumination time. Error bars represent s.d. of n = 6 spikes. (D) Effect of red laser intensity on SNR of QuasAr2 fluorescence spikes. Cells were illuminated with red laser at varying intensities and stimulated with pulses of blue light (8 pulses of 20 ms, 10 Hz, 50 mW/cm2). (E) SNR of spikes recorded in (D) as a function of red laser intensity. Error bars represent s.d. of n = 8 spikes.

DOI: http://dx.doi.org/10.7554/eLife.15202.008