Three-photon
fluorescence microscopy (3PFM) is a promising brain
research tool with submicrometer spatial resolution and high imaging
depth. However, only limited materials have been developed for 3PFM
owing to the rigorous requirement of the three-photon fluorescence
(3PF) process. Herein, under the guidance of a band gap engineering
strategy, CdTe/CdSe/ZnS quantum dots (QDs) emitting in the near-infrared
window are designed for constructing 3PF probes. The formation of
type II structure significantly increased the three-photon absorption
cross section of QDs and caused the delocalization of electron–hole
wave functions. The time-resolved transient absorption spectroscopy
confirmed that the decay of biexcitons was significantly suppressed
due to the appropriate band gap alignment, which further enhanced
the 3PF efficiency of QDs. By utilizing QD-based 3PF probes, high-resolution
3PFM imaging of cerebral vasculature was realized excited by a 1600
nm femtosecond laser, indicating the possibility of deep brain imaging
with these 3PF probes.