Journal
BIOMEDICAL MICRODEVICES
Volume 14, Issue 4, Pages 791-797Publisher
SPRINGER
DOI: 10.1007/s10544-012-9656-5
Keywords
Total internal reflection fluorescent microscopy (TIRFM); Cell analysis; Microfluidics
Funding
- Core Research for Evolutional Science & Technology (CREST), Japan Science and Technology Agency (JST), Japan
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A microfluidic device integrated with a Total Internal Reflection (TIR)-based chip for cell observation and analysis was developed. This integrated device enables in situ Total Internal Reflection Fluorescence Microscopy (TIRFM) on adherent cells cultured under continuous medium perfusion. This TIR-based chip, allows TIRFM to be easily performed on cells without the assembly of complicated optical components and cell culture chambers. The integrated device was evaluated by tracking the movement of fluorescent beads and monitoring the location of insulin granules in mouse pancreatic beta-cells. This system offers higher signal-to-noise (S/N) ratio than epi-fluorescence microscopy (EPIFM), and comparable image quality to commercial TIRFM systems when imaging insulin granules. We also detected repetitive changes in intracellular Ca2+ concentration in MIN6-m9 cells stimulated with KCl, which demonstrates quick perfusion for cell analysis while maintaining high S/N ratio.
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