Rapid multi-reagents exchange TIRFM microfluidic system for single biomolecular imaging

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Abstract A rapid multi-reagents switching microvalve system integrated with a total internal reflection fluorescence microscopy (TIRFM) was developed for real time imaging of a single protein behavior. The binding and dissociation process between a chaperonin GroEL and cochaperonin GroES was observed in this TIRFM microfluidic system. This TIRFM microfluidic system was constructed by a smooth glass in microchannel surface, that is essential for TIR imaging, and the polydimethylesiloxane (PDMS) microvalves that is operatable with a rapid response of around 100 ms. The smooth glass surface was etched with an optimized hydrofluoric acid and nitric acid (HF–HNO3) solution and had the average surface roughness of 3.2 nm. The microvalves were based on a pneumatic PDMS membrane that is beneficial for a rapid (

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