Interdisciplinary Institute of Neuroscience and Technology (ZIINT), Department of Anesthesiology, The Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, 310020, China; MOE Frontier Science Center for Brain Research and Brain Machine Integration, Zhejiang University, Hangzhou, 310058, China; State Key Laboratory of Modern Optical Instrumentations, Centre for Optical and Electromagnetic Research, College of Optical Science and Engineering, International Research Center for Advanced Photonics, Zhejiang University, Hangzhou, 310058, China.
Interdisciplinary Institute of Neuroscience and Technology (ZIINT), Department of Anesthesiology, The Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, 310020, China; MOE Frontier Science Center for Brain Research and Brain Machine Integration, Zhejiang University, Hangzhou, 310058, China.
Biomaterials. 2022 Oct;289:121809. doi: 10.1016/j.biomaterials.2022.121809. Epub 2022 Sep 16.
Multiphoton microscopy has been a powerful tool in brain research, three-photon fluorescence microscopy is increasingly becoming an emerging technique for neurological research of the cortex in depth. Nonhuman primates play important roles in the study of brain science because of their neural and vascular similarity to humans. However, there are few research results of three-photon fluorescence microscopy on the brain of nonhuman primates due to the lack of optimized imaging systems and excellent fluorescent probes. Here we introduced a bright aggregation-induced emission (AIE) probe with excellent three-photon fluorescence efficiency as well as facile synthesis process and we validated its biocompatibility in the macaque monkey. We achieved a large-depth vascular imaging of approximately 1 mm in the cerebral cortex of macaque monkey with our lab-modified three-photon fluorescence microscopy system and the AIE probe. Functional measurement of blood velocity in deep cortex capillaries was also performed. Furthermore, the comparison of cortical deep vascular structure parameters across species was presented on the monkey and mouse cortex. This work is the first in vivo three-photon fluorescence microscopic imaging research on the macaque monkey cortex reaching the imaging depth of ∼1 mm with the bright AIE probe. The results demonstrate the potential of three-photon microscopy as primate-compatible method for imaging fine vascular networks and will advance our understanding of vascular function in normal and disease in humans.
多光子显微镜在大脑研究中是一种强大的工具,三光子荧光显微镜越来越成为皮层神经科学研究的一种新兴技术。非人灵长类动物由于其与人类相似的神经和血管,在脑科学研究中发挥着重要作用。然而,由于缺乏优化的成像系统和优秀的荧光探针,三光子荧光显微镜在非人类灵长类动物大脑上的研究结果很少。在这里,我们介绍了一种具有优异的三光子荧光效率、简便的合成工艺的明亮聚集诱导发射(AIE)探针,并在猕猴中验证了其生物相容性。我们利用实验室改进的三光子荧光显微镜系统和 AIE 探针,实现了对猕猴大脑皮层约 1mm 深度的大深度血管成像。还对深层皮层毛细血管中的血流速度进行了功能测量。此外,还对猴脑和鼠脑的皮层深层血管结构参数进行了比较。这项工作是首次利用明亮的 AIE 探针在活体猕猴大脑皮层上进行的三光子荧光显微镜成像研究,达到了约 1mm 的成像深度。结果表明,三光子显微镜作为一种与灵长类动物兼容的方法,具有成像精细血管网络的潜力,并将促进我们对人类正常和疾病状态下血管功能的理解。