A major challenge for the application of strain engineering to enhance the performance of electronic devices is the quantification of strain on the nanoscale. Besides other techniques (Raman spectroscopy, X‐ray diffraction) electron beam techniques allow strain analyses with a spatial resolution of a few nanometers and a reasonable strain sensitivity of 1 × 10–3 (relative to the lattice constant of silicon). In the present work, we address practical issues in the application of nano‐beam electron diffraction (NBED) to probe the strain in strained silicon layers and sub‐100 nm structures. The investigated specimens were prepared on biaxially tensile strained silicon‐on‐insulator substrates with an initial strain of ε = 0.6% or 0.8%. Results of the NBED experiments were compared to data obtained by other strain measurement techniques; amongst them the strain mapping by peak‐pairs analysis of high‐angle annular dark field (HAADF) images was especially considered (© 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)