Fig. 3

Simulated fluorescence images permit quantification of the accuracy of cell-geometry measurements. a Simulated fluorescence images of uniform surface labeling of an in silico cell with width 2r = 1 μm and length 4 μm. The focal plane of each image relative to the cell midplane is indicated. Scale bar: 5 μm. b The measured width (magenta) of the middle cell in (a) at different offsets relative to the midplane, does not vary as strongly as the actual width (black) at that focal plane. c-e BlurLab permits the precise quantification of errors in geometry measurements from extracted contours and meshlines (c), width profiles (d), and curvature profiles (e) for cells of different radii r. Scale bar in (c): 5 μm. The colors of the meshlines in (c) are maintained in (d, e). Dashed curves in (d, e) are the actual values for the in silico cells. (f, g) Width (f) and area (g) display a systematic bias for narrow cells relative to the lines of equal measured and actual area (black). This bias is negligible for cells with width above ~1.5 μm. h Conceptual flow chart of the utility of BlurLab for quantitative comparison of experimental and simulated images to test the validity of an underlying model (here, measurements of cell size and geometry)