Fig. 2: Intracellular crystallization of MgHEX-1 in High Five insect cells and negative fluorescence (FL) imaging of the in cellulo crystals.
From: Single-cell structural biology with intracellular electron crystallography
a A median intensity Z-projection of bright-field imaging Z-stacks shows the cell suspension used for ED and serial X-ray crystallography experiments. Hexagonal-shaped MgHEX-1 crystals are indicated by black arrows. b A median intensity Z-projection of a bright-field Z-stack of a smaller region of the cell suspension, with numbers indicating individual crystals. c 3D model visualization of the region in (b) based on a confocal FL microscopy imaging Z-stack acquired using 488 nm laser excitation at room temperature. The FL excitation of enhanced yellow fluorescent protein (EYFP), co-expressed with MgHEX-1, allows for the detection of the in cellulo crystals as “sharp-edged” areas lacking the FL signal. The crystals are blue-colored for enhanced visualization. The FL virtual Z-slices in (d) and (e) show areas corresponding to the MgHEX-1 crystals labelled by numbers in (b) and lacking the EYFP FL signal. Representative images of three independent experiments are shown.
