This newly-revised course focuses on the current state-of-the-art biophysical methods that are being applied to study the structure and function of biological macromolecules and biological systems at the atomic level. We focus on all of the most informative methods, such as X-ray crystallography, Electron Microscopy and Tomography, NMR spectroscopy, mass spectrometry, and single molecule techniques. In addition, we will extensively discuss the use of combinations of methods, so called hybrid methods, including the combined use of computational tools with experimental data, to further structural elucidation of challenging macromolecular complexes and biological systems. In each section of the course, the theoretical underpinnings and the practical applications are covered. Each technique section will contain a case study, in which the students will present and discuss current research articles in class in the presence of experts in the field. The course is designed to provide a basic familiarity with the most common techniques used in structural biology and their applications to challenging problems in biology.