Yu Wentao, Zhang Yan, Lo Claudia T K, Kang Lei, Wong Terence T W
Translational and Advanced Bioimaging Laboratory, Department of Chemical and Biological Engineering, The Hong Kong University of Science and Technology, Kowloon, Hong Kong, China.
Npj Imaging. 2024 Sep 30;2(1):37. doi: 10.1038/s44303-024-00040-4.
Three-dimensional (3D) high-resolution large-volume imaging has remained a challenge. Translational rapid ultraviolet-excited sectioning tomography (TRUST) achieves rapid and cost-effective whole-organ subcellular imaging through iterative optical scanning and mechanical sectioning. However, the axial resolution is limited by the mechanical sectioning thickness or the UV light penetration depth in tissue. Here, assisted with high-and-low-frequency (HiLo) microscopy (HiLoTRUST), the optical sectioning thickness has been reduced from tens of micrometers to ~5.8 µm. In addition, HiLoTRUST has attained a finer mechanical sectioning thickness (10-15 µm) compared to TRUST (50 µm). For high-content imaging as in TRUST, we employed two additional UV light-emitting diodes (LEDs) specifically for uniform illumination. The full-color imaging ability and improved axial resolution of HiLoTRUST have been validated by two-dimensional (2D)/3D imaging of various mouse organs and human lung cancer specimens. HiLoTRUST offers a cost-effective, full-color, and high-resolution 3D imaging approach, showing its great potential in 3D histology applications.
三维（3D）高分辨率大体积成像一直是一项挑战。平移式快速紫外激发断层扫描（TRUST）通过迭代光学扫描和机械切片实现快速且经济高效的全器官亚细胞成像。然而，轴向分辨率受机械切片厚度或紫外线在组织中的穿透深度限制。在此，借助高-低频（HiLo）显微镜（HiLoTRUST），光学切片厚度已从几十微米减小至约5.8微米。此外，与TRUST（50微米）相比，HiLoTRUST获得了更精细的机械切片厚度（10-15微米）。对于TRUST中的高内涵成像，我们专门采用了另外两个紫外发光二极管（LED）用于均匀照明。HiLoTRUST的全彩色成像能力和改进的轴向分辨率已通过对各种小鼠器官和人肺癌标本的二维（2D）/3D成像得到验证。HiLoTRUST提供了一种经济高效、全彩色且高分辨率的3D成像方法，在3D组织学应用中显示出巨大潜力。
