HIGHLIGHTS
SUMMARY
Overall workflow for high‑efficiency milling of large tissue samples. The initial thickness for cryoFIB determines the milling time, which increases significantly with the initial thickness; 0.5-1.5 h are required for samples thinner than 10 μm19,20, and several hours9,21 or even days22 may be required for multicellular organisms that are tens of microns thick. The ion beam current is a key factor that determines the milling speed. The first step is the pre-sectioning, which uses a mechanical method to slice the initial sample to a workable thickness for . . .
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