Handling difficult cryo-ET samples: A study with primary neurons from Drosophila melanogaster.

ABSTRACT: Cellular neurobiology has benefited from recent advances in the field of cryo-electron tomography (cryo-ET). Numerous structural and ultrastructural insights have been obtained from plunge-frozen primary neurons cultured on electron microscopy grids. With most primary neurons been derived from rodent sources, we sought to expand the breadth of sample availability by using primary neurons derived from 3^rd instar Drosophila melanogaster larval brains. Ultrastructural abnormalities were encountered while establishing this model system for cryo-ET, which were exemplified by excessive membrane blebbing and cellular fragmentation. To optimize neuronal samples, we integrated substrate selection, micropatterning, montage data collection, and chemical fixation. Efforts to address

SUBMITTER: Kim JY 

PROVIDER: S-EPMC10369871 | biostudies-literature | 2023 Jul

REPOSITORIES: biostudies-literature