A portable fluorescence optical detection system was developed to demonstrate real-time in situ analysis of cells that are three-dimensionally cultured in an extracellular matrix under microfluidic environment. The system was designed to provide a large field of view in the lateral plane to average out cellular processes in an axial layer and simultaneously diffraction-limited axial resolution. In this proof-of-concept study, the detection system was applied to quantitative analyses of short-term measurements of cell staining and cell cytotoxicity and long-term monitoring of a cell-invasion assay. For assays, colon cancer cells were cultured in a Matrigel or alginate matrix. The measured data were largely consistent with predicted results and revealed quantitatively cell dynamics specific to 3D cell cultures. The detection system has a potential as a single package to investigate 3D cultures in a microfluidic system.
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