Fluorescence eXclusion Measurement of volume in live cells

Volume is a basic physical property of cells; however, it has been poorly investigated in cell biology so far, mostly because it is difficult to measure it precisely. Recently, large efforts were made to experimentally measure mammalian cell size and used mass, density, or volume as proxies for cell size. Here, we describe a method enabling cell volume measurements for single living cells. The method is based on the principle of fluorescent dye exclusion and can be easily implemented in cell biology laboratories. As this method is very versatile, it can be used for cells of different sizes, adherent or growing in suspension, over several cell cycles and is independent of cell shape changes. The method is also compatible with traditional cell biology tools such as epifluorescence imaging or drug treatments.