Fig. 1

Schematic illustration of (a) Principle of imaging membrane order phases in zebrafish larvae by multiphoton with Laurdan dye. Titanium-Sapphire laser produces 800 nm wavelength, and the laser beam passes through a scanner unite, scan lens (SL), tube lens (TL) all focused on the zebrafish with × 63 1.4 numerical aperture (NA) of the water-immersion lens. The scattered signals from the sample focused on two photomultiplier tube detectors (PMT) with two wavelengths; 400–460 nm (for order phase) and 470–530 nm (for disorder phase) for PMT1 and PMT2, respectively. b Laurdan Fluorescence characteristics. The Laurdan dye is excited at 800 nm and its emission wavelength peaks at ~ 450 nm (violet) when existing in the ordered phase, and ~ 500 nm in the disordered phase (blue). The violet and blue boxes represent the acquisition channels conducted in the 400–460 nm and 470–530 nm wavelength bands, respectively