Fig. 6 ∣. VAMPIRE analysis of mouse embryonic fibroblasts seeded on adhesive micro-patterned surfaces.

a, Fluorescence microscopy images of wild-type (LMNA^+/+) and lamin-deficient (LMNA^−/−) mouse embryonic fibroblasts cultured on circular (top row) and triangular (middle row) adhesive fibronectin-coated micropatterns⁷². Control cells (bottom row) are placed on the fibronectin-coated glass. Cells were fixed and stained for F-actin using Alexa Fluor 488 Phalloidin (red) and nuclear DNA using DAPI (blue). Segmented fluorescence images (right). On the left are the raw images of cells and their nuclei with the segmented contours highlighted in yellow; on the right are the same cells color coded according to the shape mode to which they belong. Scale bar, 100 μm. Inserts are magnified views of cells; scale bar, 50 μm. The identified shape modes are located on the right of the panel. b, The table on the left shows the frequency of cells classified within each shape mode for LMNA^+/+ and LMNA^−/− cells cultured on circular or triangular micropatterns (top and middle rows) and unpatterned surfaces (bottom row). The table on the right displays the values for traditional morphological parameters, including average area, shape factor (SF), and aspect ratio (AR) of cells, as well as the number of cells analyzed (#), lamin A/C status and the Shannon entropy of the cells. These results indicate that traditional morphological parameters insufficiently discriminate between the nuclear morphological responses of LMNA^+/+ and LMNA^−/− on different adhesive micropatterns (right table). By contrast, the differential morphological response of these cells is readily revealed when measured by shape mode distributions (left color-coded table). The reported values for each condition are the average abundance of cells based on two replicates of the same condition.