Endogenous Two-Photon Excited Fluorescence Provides Label-Free Visualization of the Inflammatory Response in the Rodent Spinal Cord

Figure 4

Comparison of endogenous TPEF and microglial markers in a mouse model of experimental autoimmune encephalomyelitis. (a, b) Multiphoton image of an unlabeled cryosection of a control (a) and 17 d EAE (b) mouse spinal cord obtained by combining SHG (blue channel), TPEF (green channel), and CARS (red channel). (c, d) Magnification of the area indicated by the dashed box in (b). (c) CARS channel displayed in gray scale, showing the demyelination of the tissue inside the lesion. (d) TPEF channel displayed in gray scale. (e, f) Immunohistochemistry was performed on consecutive sections. (e) Overlay of DAPI staining (blue channel) and Iba1 (red channel). (f) Overlay of DAPI staining (blue channel) and CD68 (green channel). The dotted line indicates the border of an EAE lesion. Scale bar in (a, b): 500 μm; scale bar in (c–f): 100 μm.

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