Advancements in digital microfluidic biochip(DMFB) technologies are paving the way for low-cost and automated platforms for implementing bio-protocols. However,the deployment of DMFBs outside of controlled settings will make them vulnerable to intellectual property (IP) theft. Bio-protocol development requires large investments for cross-domain innovations in biochemical analysis, microfluidics, and cyber physical systems. We propose a watermarking technique for bio-protocol IP protection – a first in microfluidics – that embeds a secret signature hierarchically across these domains. Such a signature can be attributed exclusively to the owner (like a hash).The proposed solution takes into account the inherent variability in domain-specific parameters such as mixing ratio, sensor calibration, and incubation time. We describe watermarking techniques of varying complexities for different bio-protocol steps. These include watermarking for bio-protocol synthesis parameters and the cyber physical systems control path parameters.A watermarking scheme based on integer linear programming is proposed for the sample-preparation step of a bio-protocol. The practicality of our solution is demonstrated through case studies involving an immunoassay and several mixing ratios required in the sample-preparation process of bio-protocols. The effectiveness of this approach is evaluated through various security metrics:proof of ownership score, the probability of successful tampering of the watermark, and the probability of coincidence. We also analyze the integrity of the watermark against various possible attacks: brute force search, the insertion of a new watermark,and the watermarking of more parameters.
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