Step height standards based on self-assembly for 3D metrology of biological samples

Modern microscopes and profilometers such as the coherence scanning interferometer (CSI) approach sub-nm precision in height measurements. Transfer standards at all measured size scales are needed to guarantee traceability at any scale and utilize the full potential of these instruments, but transfer standards with similar characteristics upon reflection to those of the measured samples are preferred. This is currently not the case for samples featuring dimensions of less than 10 nm and for biosamples with different optical charasteristics to silicon, silica or metals. To address the need for 3D images of biosamples with traceable dimensions, we introduce a transfer standard with dimensions guaranteed by natural self-assembly and a material that is optically similar to that in typical biosamples. We test the functionality of these transfer standards by first calibrating them using an atomic force microscope (AFM) and then using them to calibrate a CSI. We investigate whether a good enough calibration accuracy can be reached to enable a useful calibration of the CSI system. The result is that the calibration uncertainty is only marginally increased due to the sample.