First Author | Hontani Y | Year | 2021 |
Journal | Sci Adv | Volume | 7 |
Issue | 12 | PubMed ID | 33731355 |
Mgi Jnum | J:316632 | Mgi Id | MGI:6807612 |
Doi | 10.1126/sciadv.abf3531 | Citation | Hontani Y, et al. (2021) Multicolor three-photon fluorescence imaging with single-wavelength excitation deep in mouse brain. Sci Adv 7(12) |
abstractText | Multiphoton fluorescence microscopy is a powerful technique for deep-tissue observation of living cells. In particular, three-photon microscopy is highly beneficial for deep-tissue imaging because of the long excitation wavelength and the high nonlinear confinement in living tissues. Because of the large spectral separation of fluorophores of different color, multicolor three-photon imaging typically requires multiple excitation wavelengths. Here, we report a new three-photon excitation scheme: excitation to a higher-energy electronic excited state instead of the conventional excitation to the lowest-energy excited state, enabling multicolor three-photon fluorescence imaging with deep-tissue penetration in the living mouse brain using single-wavelength excitation. We further demonstrate that our excitation method results in >/=10-fold signal enhancement for some of the common red fluorescent molecules. The multicolor imaging capability and the possibility of enhanced three-photon excitation cross section will open new opportunities for life science applications of three-photon microscopy. |