Video 3.
Animated MIPs tilt series of the cell in position 3 of the crypt in Fig. 1 A. Depicted in Fig. 1 (C′ and C′′). (green) F-actin; (blue) chromatin; (red) a red line connecting the γ-tubulin–labeled spindle poles. Viewing 3D rotations of the F-actin signal combined with such a line allows visually determining spindle axis orientation with respect to crypt long axis orientation and the cleavage plane orientation with respect to the apical–basal cell polarity. Fast confocal microscopy was used to collect image stacks. An acquisition typically consisted of 60 planes × 4 channels with a z step of 0.5 µm and a pixel size of 141 nm. For creating 3D rotations of a volume comprising this cell, an image stack of a volume encompassing it was created using the Imaris 3D cropping function. Using the 3D measuring points function, a red line connecting the γ-tubulin–labeled spindle poles was incorporated into the stack.