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Simultaneous Correlative Scanning Electron and High-NA Fluorescence Microscopy

Figure 2

SCLEM of whole uncoated cells.

(a) FM image of three adenocarcinoma cells actin labeled with Alexa488. The three cells are connected via tentacles and larger extrusions. Scalebar 5 µm. (b) SEM image of the boxed area in (a), showing detailed information on the connections between the cells. A dense network of tentacles and lamellae stretches between the upper and the right cell. Scalebar 3 µm(c) FM image of an extension connecting another two adenocarcinoma cells. Clear variations in actin concentration along the extrusion can be observed. (b) BSE image of the extrusion in (a). Red arrows mark areas with increased concentration of tentacles that occur before and after the thinner parts of the extrusion. Scale bar is 10 µm. (d, e) SE and BSE high-magnification images of the boxed areas in (b) showing a region rich in tentacles and small lamellar extrusions. Scale bars are 2 µm. (f) Fluorescence intensity profiles, normalized on the maximum, taken along the red and blue lines in (a). (g) Normalized SE intensity profile taken at the corresponding locations marked in (c).

Figure 2