2025年8月21日出版的《自然—方法学》杂志发表了北京大学程和平团队的最新成果，他们提出了多功能微型双光子显微镜，可实现多色脑深部成像。

这里小组提出了FHIRM-TPM 3.0，一个2.6G微型双光子显微镜，可对行为自由的小鼠进行多色脑深部成像。该系统集成了宽带抗谐振空心光纤，传输损耗低、最低色散（700 纳米至1060 nm和激光功率的高容限）。通过校正色差和球差以及优化荧光采集孔径，课题组人员实现了深度超过820μm的皮质神经元成像 μm，并使用GRIN透镜，以单树突棘分辨率进行海马Ca²⁺成像。

此外，研究小组设计了三个可互换的共焦物镜，允许10倍的可扩展视野，最大可达1 × 0.8Mm²，横向分辨率范围为0.68~ 1.46 μm。在激发波长780nm, 920 Nm和1030 该研究团队研究了清醒的APP/PS1转基因小鼠皮层中与淀粉样斑块沉积相关的线粒体和细胞质Ca²⁺活性。因此，FHIRM-TPM 3.0提供了一个适用于各种脑成像场景的多功能成像系统。

附：英文原文

Title: A versatile miniature two-photon microscope enabling multicolor deep-brain imaging

Author: Wu, Runlong, Zhao, Chunzhu, Qiu, Shan, Zhu, Yufei, Zhang, Lifeng, Fu, Qiang, Hu, Yanhui, Wu, Dakun, Yu, Fei, Zhou, Fangxu, Huang, Haipeng, Zhang, Yunfeng, Wang, Xianhua, Wang, Aimin, Cheng, Heping

Issue&Volume: 2025-08-21

Abstract: Here we present FHIRM-TPM 3.0, a 2.6g miniature two-photon microscope capable of multicolor deep-brain imaging in freely behaving mice. The system was integrated with a broadband anti-resonant hollow-core fiber featuring low transmission loss, minimal dispersion from 700nm to 1,060nm and high tolerance of laser power. By correcting chromatic and spherical aberrations and optimizing the fluorescence collection aperture, we achieved cortical neuronal imaging at depths exceeding 820μm and, using a GRIN lens, hippocampal Ca2+ imaging at single dendritic spine resolution. Moreover, we engineered three interchangeable parfocal objectives, allowing for a tenfold scalable field of view up to 1×0.8mm2, with lateral resolutions ranging from 0.68μm to 1.46μm. By multicolor imaging at excitation wavelengths of 780nm, 920nm and 1,030nm, we investigated mitochondrial and cytosolic Ca2+ activities relative to the deposition of amyloid plaques in the cortex of awake APP/PS1 transgenic mice. Thus, FHIRM-TPM 3.0 provides a versatile imaging system suitable for diverse brain imaging scenarios.

DOI: 10.1038/s41592-025-02780-6

Source: https://www.nature.com/articles/s41592-025-02780-6