Nanotechnology has seen rapid progress in recent years, with the emergence of advanced capabilities to synthesize and characterize precisely engineered materials that point toward disruptive new performance regimes of relevance for diverse application areas. Understanding how the atomic, electronic, mechanical, and magnetic structures/properties of materials relate to their performance across multiple length-scales is, thus, of growing importance. This special topic titled “Spectroscopy, Scattering, and Imaging Techniques for Nanostructured Materials” focuses on understanding these fundamental processes, which occur within material systems in the atomic or nanoscopic regime, using advanced tools such as time-of-flight secondary ion mass spectrometry, scanning electron microscopy (SEM), X-ray diffraction (XRD), helium ion microscopy (HIM), atomic force microscopy, Raman thermometry, and in situ imaging techniques. This understanding is being leveraged by the scientific community to deliver new knowledge that has the potential to improve the performance of different material systems: lithium-ion battery materials, biological materials, nanostructured materials for energy applications, carbon nanofiber, nanoparticles, nanowires (NW), silicon microcantilevers, etc. The special topic brings together a wide variety of excellent contributions from the scientific community showcasing the depth and breadth in this vibrant topical area within nanotechnology. The collection of papers exemplifies how the current state-of-the-art of imaging and spectroscopic techniques provides new insights into these exciting nano and biological materials with unprecedented resolution.