FUN™ 1 is a unique two-color fluorescent viability probe for yeast and fungi. The FUN™ 1 stain passively diffuses into a variety of cell types and initially stains the cytoplasm with a diffusely distributed green fluorescence. However, in several common species of yeast and fungi, subsequent processing of the dye by live cells results in the formation of distinct vacuolar structures with compact form that exhibit a striking red fluorescence, accompanied by a reduction in the green cytoplasmic fluorescence. Formation of the intravacuolar structures requires both plasma membrane integrity and metabolic capability. Dead cells fluoresce bright yellow-green, with no discernable red structures.