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A conventional scanning electron microscope operated in transmission mode (TSEM) was used for imaging silica, gold and latex nanoparticles. Particles were applied to conventional transmission electron microscope (TEM) grids with different supporting films. A semiconductor detector capable of accomplishing both bright-field and dark-field imaging was used to record transmitted electrons. Particle diameter was determined from the images by comparing measured data with the results of corresponding Monte Carlo simulations which took into account particle and instrument properties. Measured and simulated line profiles agreed well; the method is sensitive to changes in diameter in the nano- and sub-nanometre range. It is concluded that TSEM imaging is a promising tool for dimensional characterization of nanoparticles. Necessary extensions to the technique in order to achieve traceable measurements are discussed.

Traceable and accurate reference dimensional metrology of nano-structures by scanning transmission electron microscopy (STEM) is introduced in the paper. Two methods, one based on the crystal lattice constant and the other based on the pitch of a feature pair, were applied to calibrate the TEM magnification. The threshold value, which was defined as the half-intensity of boundary materials, is suggested to extract the boundary position of features from the TEM image. Experimental investigations have demonstrated the high potential of the proposed methods. For instance, the standard deviation from ten repeated measurements of a line structure with a nominal 100 nm critical dimension (CD) reaches 1σ = 0.023 nm, about 0.02%. By intentionally introduced defocus and larger sample alignment errors, the investigation shows that these influences may reach 0.20 and 1.3 nm, respectively, indicating the importance of high-quality TEM measurements. Finally, a strategy for disseminating the destructive TEM results is introduced. Using this strategy, the CD of a reference material has been accurately determined. Its agreement over five independent TEM measurements is below 1.2 nm.

In recent years, DNA origami nanorulers for superresolution (SR) fluorescence microscopy have been developed from fundamental proof-of-principle experiments to commercially available test structures. The self-assembled nanostructures allow placing a defined number of fluorescent dye molecules in defined geometries in the nanometer range. Besides the unprecedented control over matter on the nanoscale, robust DNA origami nanorulers are reproducibly obtained in high yields. The distances between their fluorescent marks can be easily analysed yielding intermark distance histograms from many identical structures. Thus, DNA origami nanorulers have become excellent reference and training structures for superresolution microscopy. In this work, we go one step further and develop a calibration process for the measured distances between the fluorescent marks on DNA origami nanorulers. The superresolution technique DNA-PAINT is used to achieve nanometrological traceability of nanoruler distances following the guide to the expression of uncertainty in measurement (GUM). We further show two examples how these nanorulers are used to evaluate the performance of TIRF microscopes that are capable of single-molecule localization microscopy (SMLM).

Accurate and traceable measurements of critical dimension (CD) and sidewall profile of extreme ultraviolet (EUV) photomask structures using atomic force microscopes (AFMs) are introduced in this paper. An instrument complementarily applied with two kinds of AFM techniques, the CD-AFM and the tilting-AFM, has been developed. High measurement stability of the instrument is demonstrated, for instance, the long-term CD stability is better than 1 nm over 500 successive measurements over 55 h. To traceably calibrate the effective tip geometry, transmission electron microscopes-based method is applied, which uses either the silicon crystal lattice or the structure pitch value calibrated by metrological AFMs as an internal scale. Several grating patterns with different nominal CDs and line/space ratios of an EUV photomask have been measured using the developed methods. A data evaluation method with considered higher order tip effect due to the non-vertical sidewall is introduced. Detailed measurement results of a test EUV photomask, such as middle CD, left and right sidewall angle, feature height, line edge roughness and edge profiles are given. Finally, the AFM results are compared to that of a PTB EUV scatterometer. The comparison of the middle CD yields a linear relation within a spread of only about ±2 nm and an offset of the absolute values below 3 nm. For the sidewall angle, both methods yield consistent results within a range of about 2°.