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We demonstrate the fabrication of sharp nanopillars of high aspect ratio ontospecialized atomic force microscopy (AFM) microcantilevers and their use forhigh-speed AFM of DNA and nucleoproteins in liquid. The fabrication techniqueuses localized charged-particle-induced deposition with either a focused beamof helium ions or electrons in a helium ion microscope (HIM) or scanningelectron microscope (SEM). This approach enables customized growth ontodelicate substrates with nanometer-scale placement precision and in-situimaging of the final tip structures using the HIM or SEM. Tip radii of <10 nmare obtained and the underlying microcantilever remains intact. Instead of themore commonly used organic precursors employed for bio-AFM applications, we usean organometallic precursor (tungsten hexacarbonyl) resulting intungsten-containing tips. Transmission electron microscopy reveals a thin layerof carbon on the tips. Consequently, the interaction of the new tips withbiological specimens is likely very similar to that of standard carbonaceoustips, with the added benefit of robustness. A further advantage of theorganometallic tips is that compared to carbonaceous tips they better withstandUV-ozone cleaning treatments to remove residual organic contaminants betweenexperiments. Such contaminants are unavoidable during the scanning of softbiomolecules in liquid. Our tips can also be grown onto the blunted tips ofpreviously-used cantilevers, thus providing a means to recycle specializedcantilevers and restore their performance to the original manufacturerspecifications. Finally, a focused helium ion beam milling technique to reducethe tip radii and thus further improve lateral spatial resolution in the AFMscans is demonstrated.

Focused helium ion and electron beam induced deposition of organometallic tips for dynamic AFM of biomolecules in liquid