Deep-Penetrating and High-Resolution Continuous-Wave Nonlinear Microscopy Based on Homologous Dual-Emission Upconversion Adaptive Optics
Jing Yao, Zhipeng Yu, Yufeng Gao, Baoju Wang, Zhiyuan Wang, Tianting Zhong, Binxiong Pan, Huanhao Li, Hui Hui, Wei Zheng, Qiuqiang Zhan, Puxiang Lai
Abstract
High Resolution Image Download MS PowerPoint Slide Lanthanide-doped upconversion nanoparticles (UCNPs) are emerging as innovative nonlinear probes in biomedical studies, offering the unique capability to simultaneously emit both visible (VIS) and near-infrared (NIR) photons under continuous-wave (CW) NIR excitation. However, deep-tissue high-resolution imaging remains challenging due to the trade-off between VIS emission (higher resolution, limited penetration) and NIR emission (deeper penetration, lower resolution). Here we present a CW nonlinear microscopy based on homologous dual-emission upconversion adaptive optics, leveraging Tm 3+ /Yb 3+ co-doped UCNPs’ dual 455 nm/800 nm emission: the 800 nm emission for aberration measurement (guide-star) in deep tissues and the 455 nm emission for high-resolution imaging at matching depths. Using a home-built nonlinear laser scanning microscope with a 975 nm CW laser, we achieved near-diffraction-limited imaging (480 nm laterally) at a 500 μm depth in the mouse brain environment with significant optical aberrations. This strategy expands UCNPs’ applications and innovates the exploration of deep-tissue optical features.