“…Compared with traditional imageology, including ultrasound imaging (US), X-ray computed tomography (CT), magnetic resonance imaging (MRI), and positron emission tomography (PET), fluorescence imaging displays unique advantages, such as fine spatiotemporal resolution, real-time dynamics imaging, high sensitivity, and low cost. Modifying the tunable fluorophores or fluorescent materials in the biological environment could effectively achieve specific cell uptake and obtain high-resolution and low-background biological images. − Furthermore, these fluorophore-based fluorescence imaging techniques have also been widely used for drug distribution, metabolism assessment, disease treatment, and vascular bioimaging. − However, most current fluorophores are excited at short wavelengths (<900 nm). With the increased tissue penetration depth, they will suffer from serious photon scattering/absorption and high spontaneous fluorescence in vivo, resulting in the significantly compromised signal-to-background ratio (SBR), limiting their in vivo biological applications (Figure A). , Recent findings reveal that fluorescence imaging in the NIR-II window (1000–1700 nm) has lower photon absorption/scattering and deeper penetration depth in organisms, as well as minimal background autofluorescence to provide higher spatial resolution and improved SBR (Figure B,C); − thus, it has attracted significant attention for in vivo imaging.…”