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. 2022 Feb 24;16:839811. doi: 10.3389/fncel.2022.839811

FIGURE 1.

FIGURE 1

Functional calcium imaging from an in vivo antennal preparation. (A) Image of the in vivo preparation from the side view. (B) Schematic representation of the fly placed in a pipette tip with it’s excised antenna held in vertical position on a custom holder, for the detailed information see Supplementary Figure 1. (C) Image of the in vivo preparation from a top view, Scale bar = 0.5 mm. (D) Representation of the in vivo preparation from a top view, where open antenna is held within a #0 glass coverslip with support of an aluminum foil holder and a plastic ring around the antenna. A posterior slit on the plastic ring allowed fixing the arista directly on the #0 coverslip with odor-free glue. (E) Normalized fluorescence base level intensity from a fly preparation expressing GCaMP3.0 in Or22a olfactory neurons. The regions of interest (ROIs) are marked in yellow and the area used for the background subtraction (Bkgr) are marked in white, Scale bar = 10 μm. (F,G) Example of the recorded fluorescence intensity (expressed in ΔF/F0) over time from the same preparation as shown in (D). (F) The fly was first stimulated with mineral oil (MO, negative control); the stimulus duration is marked with a vertical gray bar, each ROI as in (E) is represented in gray and the mean value in black. (G) The fly was then stimulated with ethyl hexanoate (EH) at a 10–2 dilution in mineral oil; the stimulus duration is marked with a vertical gray bar, each ROI as in (E) is represented in light red and the mean value in red. (H) Pooled responses from n = 5 antennae to MO (black) and EH 10–2 (red). Traces represents mean ± SEM. (I) Intensity of the responses to MO (black) and EH 10–2 (red) calculated subtracting the fluorescence value at the moment of stimulation (7 s) from the fluorescence intensity at a given time expressed in seconds after stimulation. The response to EH is long lasting and is statistically significant until 20 s after stimulation [correspondent to time = 27 s plotted in (G)]. Paired t-tests, without multiple comparison correction, *p < 0.05, **p < 0.01, ns, not significant. Graphs represent mean ± SEM. Statistics for each test is reported in the Supplementary Table 4.