Fig. 2.

Workflow for CIASM. C. crescentus cultures with cells expressing PAmKate fusion proteins are plunge-frozen on electron microscopy grids. The grids are loaded onto a cryogenic microscope stage for single-molecule fluorescence imaging. The same grids are removed from the cryogenic stage and transported to a cryogenic electron microscope for CET. Following CET, a series of image transformations registers the fluorescence and CET data, resulting in the localizations of single molecules being overlaid with tomographic slices.