Keck Carl H C, Schmidt Elizabeth Lea, Zhao Su, Liu Zhongyu, Zhang Ling-Yi, Cui Miao, Chen Xiaoyu, Wang Chonghe, Cui Han, Brongersma Mark L, Hong Guosong
Department of Materials Science and Engineering, Stanford University, Stanford, CA, USA.
Wu Tsai Neurosciences Institute, Stanford University, Stanford, CA, USA.
Nat Protoc. 2025 May 13. doi: 10.1038/s41596-025-01187-z.
Optical imaging provides real-time visualization of tissues and cells at high spatial and temporal resolutions through techniques such as fluorescence microscopy, optical coherence tomography and photoacoustic imaging. However, overcoming light scattering, caused by mismatches in the refractive indices of tissue components such as water and lipids, still represents a major challenge, particularly when imaging through the thicker biological tissues of living animals. Despite advances in deep-tissue imaging, many optical methods struggle to achieve diffraction-limited resolution at depth or are unsuitable for use in live animals. Here we introduce a noninvasive approach to achieving transient and reversible optical transparency in live mice using absorbing dye molecules, using tartrazine as a representative example. Rooted in the fundamental physics of light-matter interactions, this approach enables reversible optical transparency in live animals and can be further applied ex vivo in freshly dissected tissues. In this Protocol, we detail the procedures for visualizing in vivo internal organs and muscle sarcomeres in the mouse abdomen and hindlimb through their respective transparency windows, showcasing a versatile approach for a variety of optical imaging applications in live animals. The entire protocol for an in vivo application can be implemented in just over 2 weeks by users with expertise in optical imaging and animal handling.
光学成像通过荧光显微镜、光学相干断层扫描和光声成像等技术,以高空间和时间分辨率提供组织和细胞的实时可视化。然而,克服由水和脂质等组织成分的折射率不匹配引起的光散射,仍然是一个重大挑战,特别是在对活体动物较厚的生物组织进行成像时。尽管在深层组织成像方面取得了进展,但许多光学方法仍难以在深度上实现衍射极限分辨率,或者不适用于活体动物。在这里,我们介绍一种使用吸收染料分子在活体小鼠中实现瞬态和可逆光学透明的非侵入性方法,以酒石黄作为代表性示例。基于光与物质相互作用的基本物理原理,这种方法能够在活体动物中实现可逆光学透明,并且可以进一步在新鲜解剖的组织中离体应用。在本实验方案中,我们详细介绍了通过小鼠腹部和后肢各自的透明窗口对体内内部器官和肌肉肌节进行可视化的程序,展示了一种适用于活体动物各种光学成像应用的通用方法。具有光学成像和动物处理专业知识的用户可以在短短2周多的时间内完成整个体内应用实验方案。