Fluorescence and circular dichroism, as analytical spectroscopic techniques, and mass spectrometry as an analytical tool to determine the molecular mass, provide important biophysical approaches in structural virology. Although they do not provide atomic, or near-atomic, details as electron microscopy, X-ray crystallography or nuclear magnetic resonance spectroscopy can do, they do provide important insights into virus particle composition, structure, conformational stability and dynamics, assembly and maturation, and interactions with other viral and cellular biomolecules. They can be used also to investigate the molecular determinants of virus particle structure and properties, and the changes induced in them by external factors. In this chapter, I describe the physical bases of these three techniques, and some examples on how they have helped us to understand virus particle structure and physicochemical properties.