Controlling Cross-Pollination of Biosensing Probes in Nanoscale Multiplexed Arrays

Multi-target biosensing platforms require immobilization of sensing probes on surfaces with nanoscale precision. A critical challenge is preventing unwanted molecular transfer (cross-pollination) between different probe patterns, which compromises measurement accuracy. This study investigates cross-pollination in multiplexed biosensing arrays fabricated using thermal scanning probe lithography (tSPL), a method that offers sub-10 nm spatial resolution. By systematically optimizing both the tSPL patterning parameters and chemical functionalization protocols, particularly biomolecule incubation times, this work demonstrates the ability to reduce cross-pollination to undetectable levels (0%). These experimental results establish a clear relationship between processing conditions and cross-contamination rates, providing a practical framework for reliable nanoscale patterning of multiple biomolecular probes on a single surface. These findings enable the development of high-fidelity, multi-target biosensing platforms with enhanced specificity, and therefore sensitivity, for applications in medical diagnostics, environmental monitoring, and biological research.