Video 5.
Animated MIP tilt series of a metaphase at position 5 after positioning it upright shows LOBA of a BP. (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 the apical–basal cell polarity and crypt long axis orientation. 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. Using Imaris software, image stacks were processed for 3D mark positioning. 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.