A perfusable microfluidic device with on-chip total internal reflection fluorescence microscopy (TIRFM) for in situ and real-time monitoring of live cells

Biomedical Microdevices
Ryuji YokokawaHidetoshi Kotera

Abstract

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 β-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 Ca(2+) 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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Citations

Jan 9, 2018·Chemical Society Reviews·Hui Yang, Martin A M Gijs
Mar 14, 2021·Biosensors & Bioelectronics·Shih-Mo YangYubao Cui
Dec 24, 2020·Biosensors·Yung-Tsan ChenJian-Jang Huang
Aug 31, 2021·RSC Chemical Biology·Jannik Bruun LarsenThomas Lars Andresen

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