Optical phase contrast and fluorescence images of 4T1E/M3 cells stained with anti-integrin β1 and anti-F-actin antibodies.

(A) Optical phase contrast image of cells double-stained with anti-integrin β1 and anti-F-actin antibodies after paraformaldehyde fixation and permeabilization (optical microscopy, 400× magnification). (B) Fluorescence image of anti-integrin β1 and FITC-immunostained cells under the optical microscope with a green fluorescence filter (400× magnification). (C) Fluorescence image stained with anti-F-actin antibody and rhodamine under the optical microscope with a red fluorescence filter (400× magnification). (D) Merged fluorescence image of the cells stained with anti-integrin β1 (B) and anti-F-actin antibodies (C). (E) and (F) Enlarged images of the areas enclosed by the white box in (B) and (C), respectively. (G) Merged fluorescence image of (E) and (F). (H) Enlarged image of an integrin β1 spot, indicated by the white arrow in (E). (I) Enlarged image of F-actin, indicated by the white arrow in (F). (J) Merged fluorescence image of integrin β1 (H) and F-actin (I). Scale bars: 10 μm in (A–D), 2 μm in (E–G), and 500 nm in (H–J).