Atomic force microscopy with carbon nanotube probe resolves the subunit organization of protein complexes

Among many scanning probe microscopies, atomic force microscopy (AFM) is a useful technique to analyse the structure of biological materials because o f its applicability to non-conductors in physiological conditions with high resolution. However, the resolution has been limited to an inherent proper ty of the technique; tip effect associated with a large radius of the scann ing probe. To overcome this problem, we developed a carbon nanotube probe b y attaching a carbon nanotube to a conventional scanning probe under a well -controlled process. Because of the constant and small radius of the tip (2 .5-10 nm) and the high aspect ratio (1 : 100) of the carbon nanotube, the l ateral resolution has been much improved judging from the apparent widths o f DNA and nucleosomes. The carbon nanotube probes also possessed a higher d urability than the conventional probes. We further evaluated the quality of carbon nanotube probes by three parameters to find out the best condition for AFM imaging: the angle to the tip axis; the length; and the tight fixat ion to the conventional tip. These carbon nanotube probes, with high vertic al resolution, enabled us to clearly visualize the subunit organization of multi-subunit proteins and to propose structural models for proliferating c ell nuclear antigen and replication factor C. This success in the applicati on of carbon nanotube probes provides the current AFM technology with an ad ditional power for the analyses of the detailed structure of biological mat erials and the relationship between the structure and function of proteins.