Abstract
Advanced nucleic acid amplification techniques require separating test samples into numerous units (digitalization), which enables higher sensitivity and accuracy than traditional methods. Reagent partition tools are commercially available, but are usually complex and expensive, hindering mass adoption. Herein, the fabrication of a large-scale micro-through-hole array on glass substrates for an advanced digital nucleic acid amplification technique (NAAT) is demonstrated. To meet the requirement of hole quantities (over tens of thousands) for valid nucleic acid statistics, ultrafast Bessel pulses with continuous translation of the glass substrate are applied. A single-shot Bessel pulse can lead to a single hole, and 100 holes can be produced per second (scalable to thousands per second). Experiments using the fabricated chips show that the performance meets the NAAT requirements. This work provides a highly efficient and low-cost scheme for reagent partitioning that promotes the wide accessibility of advanced digital NAATs as well as a variety of other applications.