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Cells were monitored for 8 h, and multiple image z-stacks were captured every 5 min.
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Image z-stacks were acquired in 200 nm steps.
Figure 5(c) shows image projections from a 3D rendering of the WLR image z stack.
For Extended Focal Imaging multiple Z-stacks were obtained and in-focus areas merged in Adobe Photoshop®.
For wing seta counts, wings were re-imaged at 20X through multiple focal planes (z-stacks).
Morphological reconstructions were produced manually from multiple image stacks.
Images and Z-stacks were acquired with a Nikon A1 Upright Confocal microscope.
Images and z-stacks were recorded sequentially.
We used a 25X objective on a microscope equipped with an Apotome to acquire multiple z-stack images (at least 5 z-sections, with ∼5 µm interval), which were collapsed into a single image; the degrees of deviation and the phenotype of radial glial processes were measured in a 250 µm2 zone within the cortical plate.
All images are composite z-stacks encompassing the entire cell.
Images shown are z-stacks of 5 planes with a spacing of 1 μm.
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