Fig. 7. Spontaneous motility of cultured neurospheres. (A) Representative still images of cultured neurospheres obtained using a live cell imaging apparatus. Images were acquired at the time points designated at the upper right corners. 0 h indicates a start of image tracking for the next 12 h, not the absolute start of the culture. Grid lines were drawn to facilitate tracking positions of neurospheres. Colored dots indicate the marks of neurosphere centers that are traced to quantify distances of neurosphere movements. Neurospheres marked with dots of the same color in the images at different time points are the identical ones. Boxed area indicates the magnified regions in (B). Scale bar indicates 100 µm. (B) Magnified images at the regions boxed in (A). Images at a 1-h time interval (designated at the upper right corners) starting at the 6-h time point are presented. Arrowheads indicate the tips of cytoplasmic protrusions that are traced to quantify lengths of changes in cytoplasmic protrusions. The colors of arrowheads are the same as the ones for the center marks of the neurospheres in (A) which the protrusions belong to. (C) An exemplary drawing to illustrate motile paths of neurosphere centers tracked for 24 hours by an image analysis software. Different colors are used to illustrate motile paths of different individual neurospheres. Scale bar indicates 100 µm. (D, E) Quantification graphs of the moving distance (D) of the tracked neurospheres and the length of changes in cytoplasmic protrusions (E) growing from the tracked neurospheres. Each data point represents the distance moved or the length changed during the 30-minute period between two successive imaging sessions at every 30 minutes.
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