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In situ direct laser writing (isDLW) strategies that facilitate the printing of three-dimensional (3D) nanostructured components directly inside of, and fully sealed to, enclosed microchannels are uniquely suited for manufacturing geometrically complex microfluidic technologies. Recent efforts have demonstrated the benefits of using micromolding and bonding protocols for isDLW; however, the reliance on polydimethylsiloxane (PDMS) leads to limited fluidic sealing (e.g., operational pressures <50–75 kPa) and poor compatibility with standard organic solvent-based developers. To bypass these issues, here we explore the use of cyclic olefin polymer (COP) as an enabling microchannel material for isDLW by investigating three fundamental classes of microfluidic systems corresponding to increasing degrees of sophistication: (i) “2.5D” functionally static fluidic barriers (10–100 μm in height), which supported …