Intra- and Intermolecular Synergistic Engineering of Aggregation-Induced Emission Luminogens to Boost Three-Photon Absorption for Through-Skull Brain Imaging.

Three-photon fluorescence microscopic (3PFM) bioimaging is a promising imaging technique for visualizing the brain in its native environment thanks to its advantages of high spatial resolution and large imaging depth. However, developing fluorophores with strong three-photon absorption (3PA) and bright emission that meets the requirements for efficient three-photon fluorescence microscopic (3PFM) bioimaging is still challenging. Herein, four bright fluorophores with aggregation-induced emission features are facilely synthesized, and their powders exhibit high quantum yields of up to 56.4%. The intramolecular engineering of luminogens endows (<i>E</i>)-2-(benzo[<i>d</i>]thiazol-2-yl)-3-(7-(diphenylamino)-9-ethyl-9<i>H</i>-carbazol-2-yl)acrylonitrile (DCBT) molecules with bright near-infrared emission and large 3PA cross sections of up to 1.57 × 10^-78 cm⁶ s² photon^-2 at 1550 nm, which is boosted by 3.6-fold to 5.61 × 10^-78 cm⁶ s² photon^-2 in DCBT dots benefiting from the extensive intermolecular interactions in molecular stacking. DCBT dots are successfully applied for 3PFM imaging of brain vasculature on mice with a removed or intact skull, providing images with high spatial resolution, and even small capillaries can be recognized below the skull. This study will inspire more insights for developing advanced multiphoton absorbing materials for biomedical applications.