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This paper reports on a rapid, lossless process for the bonding of poly(methyl methacrylate) (PMMA) substrates for use in microfluidic devices for blood tests, utilizing H2O plasma, and H2O linked rapid thermal annealing (RTA) treatments. The bonding of PMMA produced with H2O plasma linked with RTA was analyzed, and its effect on the bonding of free radicals was investigated. The PMMA surface treatment was performed at constant RF power and H2O vapor flow but changing plasma treatment times in both processes. The surface modification of the PMMA subjected to the H2O plasma treatment was studied by optical emission spectroscopy, which confirmed that the relative intensity of hydroxyl radicals and oxygen emission peaks. The surface of the PMMA after being subjected to the plasma linked with RTA treatment was also characterized. The free energy increased with the plasma treatment time. The tensile test results revealed that the strength of OH bonds on the PMMA surface increased with a high plasma treatment time of 120 s. After bonding, UV–visible transmission spectrum measurements revealed that the transparency of the microfluidic device increased. We demonstrated the bonding of a microfluidic device with plasma treatment without deformation. This method provides an ideal bonding technique for the manufacture of microfluidic devices with PMMA. This technique has more advantages than other bonding techniques. [ABSTRACT FROM AUTHOR]