We developed a label-free fluorescence lifetime imaging microscopy (FLIM) to quantify the metabolic state of tumor spheroids. We obtained label-free 3D FLIM images from different focal plane of cancer (MCF7) spheroids for 4 different spectral bands which was segregated by the custom-made spectral resolving unit. To obtain optically sectioned images, we used an optical fiber instead of a spatial filter. Statistical analyses showed that the signals acquired by FLIM—fluorescence intensity and lifetime—reflected well the metabolic state of cancer cells constituting spheroids. These results demonstrated the potential for quantitative measurement of the cellular metabolism in a label-free, non-destructive manner.
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