Supercontinuum-tailoring multicolor imaging reveals spatiotemporal dynamics of heterogeneous tumor evolution

Area of Science:

• Biomedical Imaging
• Cancer Research
• Microscopy

Background:

• Tumor heterogeneity and evolution are key drivers of cancer treatment failure.
• Understanding in vivo tumor dynamics requires imaging multiple components and subclones simultaneously.
• Current intravital imaging techniques face limitations in observing numerous targets concurrently.

Purpose of the Study:

• To develop a novel imaging technique for visualizing complex tumor microenvironments in vivo.
• To overcome the 'color barrier' in two-photon fluorescence imaging by enabling simultaneous observation of multiple fluorophores.
• To investigate the spatiotemporal dynamics of tumor evolution, including clonal competition and immune interactions.

Main Methods:

• Development and application of a supercontinuum-tailoring two-photon microscope (SCT-TPM).
• Utilizing phase modulation of fiber supercontinuum (SC) for excitation multiplexing with a single light beam.
• Simultaneous observation of nine fluorophores, including tumor subclones, immune cells, and vascular networks.

Main Results:

• Achieved simultaneous imaging of nine fluorophores, breaking the 'color barrier' in intravital two-photon fluorescence imaging.
• Visualized the spatiotemporal dynamics of six tumor subclones, immune cells, and vascular networks in vivo.
• Enabled the capture of rapid biological events with precise spatial alignment.

Conclusions:

• SCT-TPM offers a breakthrough for simultaneous multi-target intravital imaging.
• This technology facilitates the study of tumor evolution, clonal competition, and host-tumor interactions.
• SCT-TPM opens new avenues for tumor lineage tracking and mechanistic exploration in living systems.