Figure 2. Imaging and automated 3D single-cell segmentation of the pLLP.

(A) Maximum z-projection of a deconvolved 3D volume of the pLLP acquired using the LSM880 AiryScan FAST mode. (B) The same primordium shown with a semi-transparent color overlay of the corresponding single-cell segmentation. (C) Expanded view of the same primordium; individual segmented cells have been shifted apart without being rescaled or deformed, revealing their individual shapes within the collective. Note that the segmentation faithfully recapitulates the diversity of cell shapes within the pLLP, with the exception of fine protrusions. Since the protrusions of follower cells are often impossible to detect against the membranes of the cells ahead of them, we decided not to include fine protrusions in our analysis. All scale bars: 10 μm.

Figure 2—video 1. Dynamic View 3D pLLP Stack.

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Video of the stack shown in Figure 2A. The slices of the stack are scanned through from bottom to top first, then a maximum z-projection is generated from top to bottom. Scale bar: 10 µm.

Figure 2—video 2. Dynamic View of 3D pLLP Segmentation.

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Video of the single-cell segmentation shown in Figure 2B. The slices of the stack are scanned through from bottom to top first, then a maximum z-projection is generated from top to bottom. Scale bar: 10 µm.

Figure 2—video 3. Dynamic View of 3D Expanded Segmentation.

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3D video of expanded segmentation shown in Figure 2C. First, a rotation of the raw data is shown, followed by its expansion based on the single-cell segmentation. Scale bar: 10 µm.