Detecting Single Molecule Deoxyribonucleic Acid in a Cell Using a Three‐Dimensionally Integrated Nanopore

Amplification‐free genome analysis can revolutionize biology and medicine by uncovering genetic variations among individuals. Here, the authors report on a 3D‐integrated nanopore for electrolysis to in situ detection of single‐molecule DNA in a cell by ionic current measurements. It consists of a SiO2 multipore sheet and a SiNx nanopore membrane stacked vertically on a Si wafer. Single cell lysis is demonstrated by 106 V m−1‐level electrostatic field focused at the multinanopore. The intracellular molecules are then directly detected as they move through a sensing zone, wherein the authors find telegraphic current signatures reflecting folding degrees of freedom of the millimeter‐long polynucleotides threaded through the SiNx nanopore. The present device concept may enable on‐chip single‐molecule sequencing to multi‐omics analyses at a single‐cell level. Vertically stacked nanopore membranes enable single‐cell electrolysis to in situ intracellular biomolecule detections by ionic current measurements.