To visualize fluorescence signals from the deep tissue of the ear in live mice, we used an FVMPE-RS multiphoton laser scanning microscope (Olympus), equipped with Chameleon Vision II laser (Coherent). We used the excitation wavelength of 850 nm and green filter (495–540 nm) for GFP, and a red filter (575–645 nm) for red fluorescence signal. The ×25 objective (XLPLN25xWMP2) was used for this experiment. For live animal imaging experiments, we anesthetized mice with isoflurane anesthesia and located the mice on a stage heated to 37°C. The image data were further analyzed using ImageJ software.

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