High-throughput, bond-specific 3D chemical microscopy for lipid metabolism in cancer metastasis

Cancer metastasis relies heavily on altered lipid metabolism, yet the precise mechanisms linking fatty acid saturation states to metastatic progression remain poorly understood. Saturated and unsaturated fatty acids appear to differentially influence energy production, membrane composition, and signaling pathways, thereby shaping the tumor microenvironment and metastatic potential. To advance our understanding, we will develop and apply a novel, high-throughput, bond-specific three-dimensional (3D) optical microscopy method—Optically Detected Mid-IR 3D (O-MIR) microscopy—that overcomes current limitations in live-cell metabolic imaging. Existing approaches, including fluorescence imaging and Raman-based techniques, are constrained by labeling requirements, photobleaching, and low signal throughput. By integrating computational microscopy, mid-infrared (IR) spectroscopy, and click-free bio-orthogonal probes, our method will enable bond-specific, sub-micrometer resolution and rapid 3D live-cell imaging within both the mid-IR fingerprint and “cell-silent” spectral windows, facilitating direct, real-time tracking of fatty acids and their metabolic transformations. In Aim 1, we will develop and validate the O-MIR microscope for bond-selective 3D imaging. The O-MIR system will be constructed by integrating nanosecond-pulsed mid-IR photothermal imaging with intensity diffraction tomography algorithms, achieving bond-selective imaging with ~250 nm lateral and ~500 nm axial resolution, 3D imaging speeds of up to ~12 Hz for a 500 µm x 500 µm x 100 µm volume, and high mid-IR spectral fidelity. We will validate its performance using microsphere phantoms and fatty acid suspensions in both the mid-IR fingerprint and cell-silent windows. In Aim 2, we will employ the O-MIR microscope to investigate how fatty acids with differing degrees of saturation influence breast cancer cell metastasis, comparing non-metastatic and metastatic cell lines under metabolic inhibition. We will quantify changes in fatty acid uptake, localization, and metabolic processes that enhance or impede metastatic phenotypes. By clarifying how specific fatty acids modulate metastasis, this work will identify novel metabolic markers and therapeutic targets. Ultimately, our findings will inform the development of enzyme-targeting strategies and metabolic interventions to reduce metastasis and improve patient outcomes. Project Number: 5P20GM135009-05 | Fiscal Year: 2026 | NIH Institute/Center: National Institute of General Medical Sciences (NIGMS) | Principal Investigator: JIAN ZHAO | Institution: UNIVERSITY OF OKLAHOMA, NORMAN, OK | Award Amount: $155,484 | Activity Code: P20 | Study Section: ZGM1-RCB-2(C1) View on NIH RePORTER: https://reporter.nih.gov/project-details/11473632