De Novo Design of Aggregation-Induced Emission Luminogen for Three-Photon Fluorescence Imaging of Subcortical Structures Excited at Both NIR-III and NIR-IV Windows

Three-photon fluorescence (3PF) imaging excited at 1700 nm window is an enabling technology for visualizing deep brain structures and dynamics. Recently, the 2200 nm window has emerged as the longest excitation window suitable for deep-brain 3PF imaging. Bright fluorescent probes lay the material basis for deep-brain 3PF imaging. Among various fluorescent probes, aggregation-induced emission luminogens (AIEgens) have great potential in 3PF imaging excited at the 1700 nm window in vivo. However, to the best of knowledge, there is no AIEgens applicable to 3PF imaging excited at both the 1700 and 2200 nm windows. To readily fill this gap, here this study designs and synthesizes a novel AIEgen, namely TPE-DPTT-ICP, which generates bright 3PF signals excited at both 1700 and 2200 nm. The accordingly fabricated TPE-DPTT-ICP nanoparticles (NPs) possess excellent water dispersibility, colloidal stability, biocompatibility, photostability and large 3P action cross section, key to in vivo imaging. In mouse brain in vivo, TPE-DPTT-ICP NPs enable deepbrain 3PF imaging of subcortical structures excited at both the two windows, reaching depths of 1640 and 880 mu m below the brain surface, respectively. TPE-DPTT-ICP NPs are thus a versatile material simultaneously catering to the need at two infrared optical windows with deep tissue penetration.