At the Laboratory for Precise Measurements of Length, currently the Croatian National Laboratory for Length, unique nano-roughness calibration standards were developed, which have been physically implemented in cooperation with the company MikroMasch Trading OU and the Ruder Bošković Institute. In this paper, a new design for a calibration standard with two measuring surfaces is presented. One of the surfaces is for the reproduction of roughness parameters, while the other is for the

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... for the traceability of length units below 50 nm. The nominal values of the groove depths on these measuring surfaces are the same. Thus, a link between the measuring surfaces has been ensured, which makes these standards unique. Furthermore, the calibration standards available on the market are generally designed specifically for individual groups of measuring instrumentation, such as interferometric microscopes, stylus instruments, scanning electron microscopes (SEM) or scanning probe microscopes. In this paper, a new design for nano-roughness standards has been proposed for use in the calibration of optical instruments, as well as for stylus instruments, SEM, atomic force microscopes and scanning tunneling microscopes. Therefore, the development of these new nano-roughness calibration standards greatly contributes to the reproducibility of the results of groove depth measurement as well as the 2D and 3D roughness parameters obtained by various measuring methods.