WCLC 2025 - Posters & ePosters-EP.03.16 Cell-Specific Fluorescence Lifetime Values in Live Non-Small Cell Lung Cancer Cells

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This study investigates fluorescence lifetime imaging microscopy (FLIM) as a tool for characterizing metabolic differences within live non-small cell lung cancer (NSCLC) cells and tissues. FLIM measures the decay rates of endogenous autofluorescent metabolites such as NAD(P)H and FAD, providing real-time, label-free insight into cellular metabolism. Previous work showed decreased fluorescence lifetimes in NSCLC tissue compared to adjacent non-tumorous lung; however, this study focuses on dissecting the cellular and regional heterogeneity underlying these signals.<br /><br />Using single-photon FLIM at 448 nm excitation, the researchers measured intrinsic autofluorescence in 2D single-cell cultures and 3D organoids derived from tumor and non-tumor lung tissue. Results confirmed that fluorescence lifetimes vary by cell type and are conserved within each, highlighting cellular specificity. Organoids demonstrated metabolic heterogeneity with signals consistent with free FAD excitation, suggesting that FLIM can distinguish structural components based on metabolic activity in a 3D context.<br /><br />At the tissue level, fluorescence lifetime varied by region, reinforcing metabolic differences within the tumor microenvironment. An adenosine perturbation assay showed that extracellular adenosine exposure decreased mean fluorescence lifetime, supporting its influence on metabolic signaling detectable by FLIM.<br /><br />Overall, the study establishes FLIM as a powerful translational technique to differentiate NSCLC tumor cell populations and their metabolic states in real time without external labels. It underscores the method’s potential in early diagnosis and tumor characterization by linking autofluorescence decay patterns to specific cell types and microenvironmental conditions. The findings pave the way for using FLIM in assessing metabolic alterations that could inform prognosis and therapeutic targeting in NSCLC.<br /><br />Acknowledgments highlighted support from supervisors, the university’s imaging and histology facilities, and cited relevant foundational literature on FLIM and metabolism in cancer.

Asset Subtitle

Ava Russell

Meta Tag

Speaker Ava Russell

Topic Tumor Biology – Translational Biology

Keywords

fluorescence lifetime imaging microscopy

FLIM

non-small cell lung cancer

NSCLC

metabolic heterogeneity

autofluorescence

NAD(P)H

FAD

3D organoids

tumor microenvironment