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We report on design of fluorescently-labeled DNA that was used to assemble nucleosomal nanoparticles with two linkers of 40 base pair size. To study mutual conformation of nucleosomal linkers with spFRET microscopy, one linker was labeled with donor (Cy3) and another one with acceptor (Cy5) fluorophore. Designed nucleosomal nanoparticles are fluorescent sensors that are sensitive to mutual conformation of linkers. Using spFRET microscopy in combination with these sensors the important structural aspects of complexes of nucleosomes with various chromatin architectural proteins can be decoded.