Chemical responses of single yeast cells studied by fluorescence microspectroscopy under solution-flow conditions

A microspectroscopy system combined with a fluid manifold was developed to manipulate and analyze "single" living cells. A sample buffer solution containing living cells was introduced into a flow cell set on a thermostated microscope stage and a few cells were allowed to attach to the bottom wall of the flow cell. With these living cells being attached to the wall, other floating cells were pumped out by flowing a buffer solution. These procedures made it possible to keep a few cells in the flow cell and to analyze single cells by fluorescence microspectroscopy. The technique was applied to study the time course of staining processes of single living yeast (Saccharomyces cerevisiae) cells by using two types of a fluorescent probe. The present methodology was shown to be of primary importance for obtaining biochemical/physiological information on single living cells and also for studying cell-to-cell variations in several characteristics.