A method is demonstrated for large-scale micro-through-hole (MTH) array fabrication in glass substrates for an advanced digital nucleic acid amplification technique (NAAT). The fabricated chip is advantageous to current partitioning devices in terms of speed, cost, and simplicity. To satisfy the requirement of MTH quantities for valid nucleic acid statistics, we improve the laser processing speed by focusing ultrafast Bessel pulses into a glass substrate under continuous translation. A single Bessel pulse can result a single MTH, and hundreds to thousands of MTHs can be produced per second. Preliminary digital NAAT experiment shows promising results of reagent partitioning of the fabricated chip. This work offers a highly efficient and low-cost scheme for glass-based reagent partitioning that will contribute to the wide accessibility of advanced digital NAATs.
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