Abstract Nano and micro coordinate measuring machines (CMMs) have been developed for the characterization of small dimensional features. They require a procedure which enables a traceable and precise characterization of probing spheres. In this contribution we explore the use of well characterized micro spheres as reference artifacts for the in-situ characterization of probing spheres along the probing sphere’s equator. The spheres are characterized using a strategy which is based on a set of tactile surface scans in conjunction with a stitching-algorithm. These micro spheres serve as a reference for the in-situ characterization of a tactile 3D micro probe on a nano measuring machine (NMM-1). Our investigations are based on a sample of eight spheres sourced from two different suppliers. Although the sample is small, we could already observe characteristics which seem to be typical for spheres of a certain type (i.e. nominal radius and material). The experiments indicate that micro spheres are a suitable reference artifact for tactile 3D micro probes. We were able to reproduce the measured mean radius of the probing sphere with a standard deviation of 31 nm using reference spheres whose nominal radius covers a range of 89 µm (61 µm up to 150 µm).