Plasma microcontact patterning (PμCP) and replica molding were combined to make PDMS/glass microfluidic devices with β-cyclodextrin (β-CD) patterns attached covalently on the glass surface inside microchannels. The supramolecular reactivity, reusability and association constant of β-CD with Cy5–Ad₂ was tested by analyzing signal-to-noise ratios of patterns vs. spacing with fluorescence microscopy.