一种多功能微型双光子显微镜,可进行多色深层脑部成像
Runlong Wu1,2,3, Chunzhu Zhao4,5, Shan Qiu6
1National Biomedical Imaging Center, State Key Laboratory of Membrane Biology, Institute of Molecular Medicine, Peking-Tsinghua Center for Life Sciences, College of Future Technology, Peking University, Beijing, China. rlwu@bistu.edu.cn.
Nature methods
|August 21, 2025
概括
我们开发了FHIRM-TPM 3.0, 这种先进的双光子显微镜系统能够在体内对神经元活动和细胞结构进行多色成像.
科学领域:
- 神经科学
- 生物医学工程
- 显微镜技术
背景情况:
- 对于理解神经回路至关重要.
- 现有的双光子显微镜系统面临尺寸,灵活性和成像深度的限制.
- 多色成像对于解剖大脑中复杂的细胞和分子过程至关重要.
研究的目的:
- 为了介绍FHIRM-TPM 3.0, 一个新的,紧的两光子显微镜用于先进的体内脑图像.
- 展示该系统在自由行为小鼠中的多色深层脑图像的能力.
- 展示该系统的多功能性和高分辨率,
主要方法:
- 微型两光子显微镜与宽带反共振空心纤维的集成.
- 纠正光学偏差并优化光收集以深入组织.
- 设计可互换的目标以实现可扩展的视野和高侧分辨率.
- 使用多色激发波长 (780,920,1030nm) 进行同时监测细胞活动.
主要成果:
- 在超过820μm的深度实现皮层神经元成像.
- 通过使用GRIN镜头实现单一树突脊柱分辨率的海马Ca2+成像.
- 提供十倍可扩展的视野 (高达1×0.8mm2),分辨率从0.68μm到1.46μm.
- 在APP/PS1小鼠中成功研究了与粉样斑块相关的线粒体和细胞质Ca2+活动.
结论:
- FHIRM-TPM 3.0 是用于神经科学研究的多色深层脑成像的多功能且强大的工具.
- 该系统的小型化和先进的光学设计使其在自由行为对象中进行体内研究.
- 它使得高分辨率,深层组织成像, 推进神经疾病和大脑功能的研究.
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