Figure 10

A detailed analysis of the edge of a lamellipodium of an unlabeled endothelial cell. This example illustrates analysis by counts per pixel and HSI. (a-c) Three MIMS images acquired in parallel at (a) ¹²C¹⁴N^-, (b) ¹²C^-, and (c) ¹⁶O^-. Field: 60 × 60 μm (256 × 256 pixels); acquisition time 2 min. (d) HSI image of the ratio ¹²C¹⁴N^-/¹²C^- (the numerator has been multiplied by 10). Magenta dots (arrowed) indicating areas of high relative ¹⁵N incorporation appear at the edge of the lamellipodium. Field: 60 × 60 μm (256 × 256 pixels). (e) HSI images of the ratios ¹²C¹⁴N^-/¹²C^- (left) and ¹⁶O/¹²C¹⁴N^- (right; the numerator has been multiplied by 10). The regularly spaced dots (arrowed) can be seen at the edge of the lamellipodium. (f) At each pixel, arranged from top to bottom, are the values of the ¹²C¹⁴N^- counts, the ¹²C^- counts and the ¹²C¹⁴N^-/¹²C^- ratio (multiplied by 10) for the pixels shown in the inset in (e). (g) The corresponding values at each pixel, arranged from top to bottom, for the ¹²C¹⁴N^-/¹²C^- ratio values (multiplied by 10) and the ¹⁶O^-/¹²C¹⁴N^- ratio (multiplied by 10). (h,i) Bar graph of the mean count values of ¹²C^-, ¹²C¹⁴N^- and ¹⁶O^- on (h) the dot at the periphery of the lamellipodia and (i) the edge of the lamellipodia.