What are the advantages of two-photon fluorescence microscopy
1) Long-wavelength light is less affected by scattering than short-wavelength light and easily penetrates the specimen;
2) The fluorescent molecules outside the focal plane are not excited, so that more excitation light can reach the focal plane, so that the excitation light can penetrate deeper specimens;
3) Long-wavelength near-infrared light is less toxic to cells than short-wavelength light;
4) When observing specimens with a two-photon microscope, photobleaching and phototoxicity occur only in the focal plane. Therefore, two-photon microscopy is more suitable than single-photon microscopy for observing thick specimens, for observing living cells, or for spot photobleaching experiments.
