Fig. 2: Super-resolution live imaging of QL.p division and downstream image processing.

a Super-resolution live two-colour timeseries of QL.p division in animals expressing the transgenic multicopy array bcIs153. Plasma membrane (myristoylated mCherry) and chromatin (mCherry::his-24) are shown in magenta, and mitochondria (mtGFP) in cyan. Images are maximum intensity projections of aligned z-stacks (top view, xy). Image orientation has been changed for time-series illustration, whereas the original orientation can be seen in image processing (c–e). A anterior side, P posterior side, V ventral side, D dorsal side. b Lateral view (xz) of a z-stack showing QL.p at metaphase before (left) and after (right) alignment. Images refer to a cropped volume from the original z-stack (see also (d)). c Magenta z-stacks (plasma membrane + chromatin (mCherry), left) are subtracted from cyan z-stacks (‘GFP (original)’, centre). This arithmetic operation has minor effects on mtGFP signal distribution (‘GFP (subtracted)’) (see also Supplementary Fig. S3a). The subtraction is conducted on the entire z-stack. d Maximum intensity projection of an aligned and subtracted z-stack, which shows QL.ap in contact with the anterior side of QL.p at metaphase (see also Supplementary Fig. S2). The image represents the field-of-view (FOV) used to record QL.p division. The white box represents the ROI used to duplicate a new z-stack, after alignment, which will be used for the last step of image processing (e). e Richardson–Lucy (RL) deconvolution (step 4) of both magenta and cyan z-stacks. Deconvolved images are used for 3D rendering of QL.p cell shapes and mitochondria. Steps in (b–e) are performed in Fiji and describe the entire image processing workflow. c–e Refer to (a) at time −1 min.