Fig. 4

AO improves three-color intravital imaging of a tumor microenvironment. a The tumor microenvironment was imaged through a dorsal skin-fold window chamber. The tumor cells, host cells and arterial vessels were labeled with mCerulean, EGFP and Alexa Fluor 633, respectively. b Maximum intensity projection of the microvessels centered 92 μm below the tissue surface, with which the aberration was measured. c Maximum intensity projection (XY) of the image volume 76–81 μm below the tissue surface, and image of the axial section (XZ) indicated by the red arrowheads without and with AO correction. The yellow dashed boxes indicate the positions of the microvessels used for aberration measurement. d Intensity profiles along the pink, orange and cyan lines in (c). e Zoomed-in views of the white boxes in (c). The images are enhanced with a 2x digital gain to increase the visibility. The yellow arrowhead indicates a microvessel. f Zoomed-in views of the orange boxes in (c). g Corrective wavefront. The dashed squares indicate the layout of the wavefront segments, where each wavefront segment corresponds to a unit square. A 2 × 2 with 1 × 1 stepped overlapping mask approach was used in the measurement process. Scale bars, 5 μm in (b), 10 μm in (c), 4 μm in (e), and 2 μm in (f)