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Fig. 4
Ca2+-imaging. Transverse plane of a 4 dpf elavl3:H2B-GCaMP6s larva imaged with Gaussian (a) and Bessel beam illumination (b). Scale bar is 50 μm. Yellow arrow indicates direction of light sheet. Indicated are three exemplary cells with red (cyan) circles which have been illuminated with a Gaussian (Bessel) beam and whose traces can be seen in (c). (d) Mean standard deviation (STD) of traces without neuronal activity (p < 0.0001, paired t-test, n = 625 cells in N = 7, error is sem). (e) Exemplary dF/F traces measured with Bessel beam illumination during neuronal activity. Indicated are peaks (triangles) above the noise level for Gaussian (red) and Bessel beam illumination (cyan). Peaks with prominence below threshold were discarded (black arrow head). (f) Peaks per minute detected above noise level using appropriate thresholding to exclude Gaussian and Bessel baseline noise (p < 0.0001, paired t-test, n = 586, in N = 1 larva, error is sem). (g) Transverse plane of a 4dpf larva imaged with Gaussian illumination. Indicated are cells located on two adjacent excitation lines (red, blue) and randomly in the hindbrain as part of an active network (yellow, see also zoomed view in h). (i) Exemplary traces of cells (marked yellow in h) measured with Gaussian (red) and Bessel beam illumination (cyan). (j) Correlation matrix of cells measured with Gaussian (lower triangular) and Bessel beam illumination (upper triangular matrix). The color scale ranges from –1 (blue) to +1 (red). (k) Averaged coefficient of cross correlations (ccc) of yellow yellow quadrant (p < 0.0001, paired t-test, n = 91, N = 1, error is sem).