Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan, Hubei, China.
MoE Key Laboratory for Biomedical Photonics, Huazhong University of Science and Technology, Wuhan, Hubei, China.
Nat Commun. 2024 Oct 29;15(1):9313. doi: 10.1038/s41467-024-53697-1.
Tumor heterogeneity and tumor evolution contribute to cancer treatment failure. To understand how selective pressures drive heterogeneous tumor evolution, it would be useful to image multiple important components and tumor subclones in vivo. We propose a supercontinuum-tailoring two-photon microscope (SCT-TPM) and realize simultaneous observation of nine fluorophores with a single light beam, breaking through the 'color barrier' of intravital two-photon fluorescence imaging. It achieves excitation multiplexing only by modulating the phase of fiber supercontinuum (SC), allowing to capture rapid events of multiple targets with maintaining precise spatial alignment. We employ SCT-TPM to visualize the spatiotemporal dynamics of heterogeneous tumor evolution under host immune surveillance, particularly the behaviors and interactions of six tumor subclones, immune cells and vascular network, and thus infer the trajectories of tumor progression and clonal competition. SCT-TPM opens up the possibility of tumor lineage tracking and mechanism exploration in living biological systems.
肿瘤异质性和肿瘤进化导致癌症治疗失败。为了了解选择压力如何驱动异质性肿瘤进化，在体内对多个重要成分和肿瘤亚克隆进行成像将是很有用的。我们提出了一种超连续谱定制双光子显微镜（SCT-TPM），并通过单光束实现了对 9 种荧光团的同时观察，突破了活体双光子荧光成像的“颜色障碍”。它仅通过调制光纤超连续（SC）的相位来实现激发复用，从而在保持精确空间对准的同时，能够捕捉多个目标的快速事件。我们利用 SCT-TPM 可视化宿主免疫监视下异质性肿瘤进化的时空动力学，特别是六个肿瘤亚克隆、免疫细胞和血管网络的行为和相互作用，从而推断肿瘤进展和克隆竞争的轨迹。SCT-TPM 为在活体生物系统中进行肿瘤谱系追踪和机制探索开辟了可能性。
