This protocol describes the preparation and use of spherical indenters for micrometer-scale imaging and mechanical testing with atomic force microscopy (AFM). A spherical indenter is prepared by gluing a hard borosilicate sphere to a tipless cantilever. For this purpose, a stereomicroscope with a micromanipulator attachment is employed. The spheres are cleaned prior to gluing to remove any contamination from their surfaces. Once they are cleaned, the borosilicate spheres are stored in absolute ethanol in a glass tube.
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