ZFIN Image: Tanimoto et al., 2022, Fig. 1

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Figure Caption

Fig. 1

Tiltable objective microscope enables functional imaging during 360° static tilt and vibration.

a Schematic showing a tiltable objective microscope. An objective lens and a specimen are tilted by a motorized rotation stage (see Supplementary Movie ¹). b Gravitational vector components (magenta and blue arrows) during static tilt produced by large-angle, slow rotation (left). Inertial acceleration during vibration (orange arrows) produced by small-angle, fast rotation (right) (see Supplementary Fig. ¹). c Rotated images are merged and registered by correcting the image rotation (see Supplementary Movie ² and “Methods”). d Fluorescent beads. Images from 3 fields of view (FOVs) are superimposed. Scale bar: 20 μm. e Time course of beads fluorescent intensity changes during 360° tilt. Large fluctuation in green and red signals is reduced in ratio and ratio change (ΔR/R₀). 116 beads from 3 FOVs. f Relationship between tilt angle and ΔR/R₀. g Distribution of ΔR/R₀ values during tilt. h Relationship between beads distance from the center of image rotation, which is the center of the image in [d], and ΔR/R₀ maximum (red) and minimum (blue). i Relationship between the angular position of beads around the center of image rotation, which is the center of the image in [d], and ΔR/R₀ maximum (red) and minimum (blue). j Brain neurons expressing green- and red-Kaede in Tg(nefma-hs:Gal4; UAS:Kaede) larval zebrafish at 5 dpf. Images from 17 FOVs are superimposed. Red channel is shown in grayscale. Positions of neurons are labeled by FOV colors. Scale bar: 10 μm. k Time course of neuronal fluorescent intensity changes during 360° tilt. Large fluctuation in green and red signals is reduced in ratio and ΔR/R₀. 116 neurons from 17 FOVs. l Relationship between tilt angle and ΔR/R₀. m Distribution of ΔR/R₀ values during tilt. n Relationship between neuron distance from the center of image rotation, which is the center of the image in [j], and ΔR/R₀ maximum (red) and minimum (blue). i Relationship between angular position of neurons around the center of image rotation, which is the center of the image in [j], and ΔR/R₀ maximum (red) and minimum (blue). Source data are provided as a Source Data file.

Acknowledgments

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