Figure 6
3D CLEM experiment of 99.2 nm diameter fiducials after cellular uptake in HeLa cells. (a) Confocal fluorescence microscopy slice of cells after fixation. White areas indicate coarse outlines of two cells. The enlarged region highlighted in green is chosen as ROI for FIB-SEM imaging. The white arrowheads indicate fiducials visible in the corresponding FIB-SEM slice shown in (b). (b) Virtual slice of the reconstructed FIB-SEM data, corresponding to the focal plane used for fluorescence imaging. Red arrowheads indicate compartments visualized in fluorescence microscopy, which contain various amounts of fiducials. The organelles encircled in blue and purple containing fiducials are enlarged in (c). (c) FIB-SEM slices through organelles of interest at different heights of both compartments, clearly showing the different amounts of fiducials present. The numbers below the figures correspond to the number of the FIB-SEM slices. (d) Overlay of FM and EM data based on the positions of the fiducials indicated by the arrowheads in (a,b). The scale bars in (a) correspond to 20 and 2 μm, the scale bars in (b,d) correspond to 2 μm.
