Fig. 1

(a) Flowchart showing the processes from tissue slide preparation to histological analysis and two-photon image analysis. (b) The microscope setup for two-photon imaging. SU: scan unit; L: lens; M: mirror; DC: dichroic beamsplitter; OBJ: objective lens; S: sample; 3D-MS: three-dimensional motorized stage; CF: color filter; F: filter; PMT: photomultiplier tube. (c) The demonstration of a large-field image (top) tiled from single 2D images (bottom). The green‒yellow color indicates the SHG presenting collagen fibers, while the red color indicates the TPF presenting layered histopathological structures.