High‐Fidelity Dehydration and Low‐Artifact Scanning Electron Microscopy Measurement of DNA Self‐Assembled Nanopatterns

DNA self‐assembly achieves patterning critical dimension (CD) comparable to or beyond extreme ultraviolet lithography, providing a programmable bottom‐up solution to nanoscale fabrication. In this context, high‐fidelity dehydration and low‐artifact metrology of DNA nanopatterns on solid substrates are essential for advancing nanomanufacturing. Conventional scanning electron microscopy (SEM) measurements require metal sputtering to visualize 10 nm scale insulating samples, which alters the intrinsic morphology and chemical composition of raw samples. Here, we report a low‐distortion immobilization method for DNA brick‐based crystalline films enabling direct SEM imaging without metal sputtering. The incorporation of Ni 2+ ions inhibits the dehydration‐induced deformation of DNA nanopatterns on solid substrates. Moreover, Ni 2+ ions react with the DNA phosphate backbone to form semiconducting nickel phosphate derivatives, enhancing electrical and thermal conductivity. Thus, Ni 2+ ‐incorporated DNA crystalline films prevent charge accumulation and thermal deformation during accurate SEM imaging. This manipulation reduces CD measurement deviation by 50% versus conventional metal‐sputtering SEM metrology for DNA nanopatterns. The whole strategy is applicable to Si, SiO 2 , and graphene substrates. The as‐measured DNA nanopatterns will be compatible with reactive ion etching, chemical vapor deposition, and atomic layer deposition, supporting extensive DNA‐based nanomanufacturing technologies.