Fusome topology and inheritance during insect gametogenesis
Fig 2
Pipeline for 3D fusome reconstructions in developing germline cysts.
(A) Starting from 3D confocal images of fusomes with fluorescently-labeled ring canals and fusome, training in a machine learning program and processing in MATLAB as described in [44, 45] produced a 3D reconstruction of the fusome and ring canal system at successive stages of germline cyst formation (Fig 1A). From here, ring canals at the junction between sister cells were used to segment the fusome into its component parts (multicolored cells) and, in the case of oogenesis, produce an adjacency matrix representing intercellular connections within the cyst to compare with the known female germline cyst network topology, permitting unambiguous cell label assignments to the individual fusome fragments. These fusome fragments were then labeled according to their position relative to the presumptive oocyte. (B) Male cysts underwent the same procedure of collective adjacency information, but as no oocyte is present, no distinction was made between cells. Scale bars = 2 μm (A) and 10 μm (B).