Fig. 10

Imaging of subcellular dynamics. (A) Maximal projection of a Z-stack showing cells in the midbrain of a 3 dpf embryo double-transfected with pCS-mCitrine and pCS-MitoTag-RFP. Anterior is to the left. Shown is one embryo out of several successful electroporated embryos from one electroporation approach. (B) Magnified single plane from the Z-stack projection shown in (A), zoomed in. (C) Putative motorneuron in culture co-transfected with pCS-MitoTag-YFP and pCS-mCherry (1 dap). Inset: Two frames of Movie 3 visualizing the dynamics of mitochondria (arrowheads). (D) Neuron-like cell in culture (1 dap) co-transfected with pCS-MitoTag-YFP and pCS-mCherry (1 dap). Mitochondria in the cell body are highly dynamic as shown by two frames of Movie 4 displayed in (E). (F) Cultured cell (1 dap) transfected with pCS-VAMP1-mCitrine. Two frames of Movie 5 are shown. Arrowheads label the dynamic changes of cellular projections. (G) XITubb:DsRed-cell in culture (1 dap) transfected with pCS-VAMP1-mCitrine. (H) Two frames of a time lapse showing the cell body of the cell shown in (G). Arrowheads label the movement of a single vesicle. Scale bars, 10 µm. Each transfection was at least performed three times. The figure shows a collection of images obtained from several preparations.