The resurgence in the use of lightweight materials highlights the need to consider vibration in the design of floors to ensure not only the stability of the building but also the comfort of the occupants. The aim of this work was to define the most appropriate acceptance thresholds to avoid over-designing the floor while ensuring a sufficient level of comfort. Firstly, controlled laboratory tests were carried out to identify the main factors influencing perceived comfort and to define test protocols to limit bias from subjective aspects. A shaker table was used to generate vertical vibrations to which participants were exposed. Representative tests were then carried out on a wooden laboratory floor with humans as the vibration source. Field tests were then carried out in the same way as the laboratory tests in public buildings made of wood and metal. Each time the floors were instrumented, the participants were interviewed, and their physiological indicators were measured. Our main interest was assessing the comfort level of seated people in offices and housing. At relatively low levels of vibration, participants may have felt dizzy, unable to concentrate on a task or physiologically uncomfortable. A relationship between the level of acceleration and the level of discomfort has been demonstrated. All of these experiments have made it possible to define, under certain conditions, maximum acceleration levels that could lead to moderate or intolerable discomfort.