Fig. 4
The OptoArm fulfills all technical criteria in different experimental set-ups. A The OptoArm set-up used with a standard dissection microscope. B In experimental set-ups, the OptoArm is used with a specific angle (43°), thereby deviating from perpendicular illumination. C The different illumination profiles for the different lenses upon and illumination with the specified angle of incidence. D The different integrated intensity profiles for C. The histograms show the distribution of intensity readings between a minimum and maximum intensity with a specified bin width. Red lines show the average intensity. E Schematic showing the different visible microscopic areas (of the used microscope) when using different magnifications. The areas are on scale and can be directly compared to the intensity maps shown in D. 1.0× corresponds to a total magnification of 15×, and 4.0 to a total of 60×. The dotted red line represents the area in D with an intensity of > 1.6 mW/mm2. F The OptoArm set-up with the WF-NTP. G The different integrated intensity profiles of the LED with the 21° lens, at different working distances with a fixed angle of incidence of 43°. Profiles were generated in the set-up with the WF-NTP. Circles represent the outline of a 3-cm NGM plate. H Histograms of intensity readings of the microscope-light (Gaussian) and WF-NTP backlight (flat-field). Red lines show the average intensity. All readings were performed at 448 nm. I Thrashing frequency of D1 worms, expressing ChR2 under the unc-17 (acetylcholine) or unc-47 (GABA) promotor, recorded with the WF-NTP, grown with or without ATR. There is no significant effect of the WF-NTP light on this behavior, n = 15, two-tailed unpaired Student’s t test (all: n.s.). J Body length of D1 worms recorded with the WF-NTP, grown with or without ATR. Body length was normalized by the mean of the paired ATR− condition. There is no significant effect of the WF-NTP light on this behavior, n = 15, two-tailed unpaired Student’s t test (all: n.s.). For I and J, there was no photo-stimulation with the OptoArm