Supplementary Figure 12: soSPIM calibration.
From: 3D high- and super-resolution imaging using single-objective SPIM
(a) Plot showing the measurements of the axial z-position of the light sheet depending on the light-sheet displacement along the mirror x-axis for the 60× water-immersion objective (blue dots) and the 40× (red dot), 20× (red dots) and 10× (yellow dots) objectives as described in the Online Methods. The black lines represent the linear fits used as calibration for soSPIM 3D-volume imaging. (b) Plot showing the position of the thinnest part of the light sheet (x0) along the x-axis depending of the defocus strength applied to the focus-tunable lens (red dots). The black line represents the linear fit used to calibrate the defocus system of the soSPIM microscope. (c) Plot showing the ratio between the x-axis drift of the light-sheet position (dx0) per z-axis objective displacement steps (dz) depending on the defocus strength applied to the focus-tunable lens (dI) per z-axis objective displacement steps (dz) (red dots) for the 60× water-immersion objective. The black line represents the linear fit. To fully compensate for the x-drift of the light sheet for 3D imaging the ratio dx0/dz must be set to 0, i.e., dI/dz = 0.013 mA/µm.
