(620av) Sorting and Analysis of Yeast, Based on Replicative Age, Using High Reynolds Inertial Focusing (Rapid Fire)

In recent years, yeast has emerged as a valuable model in the study of aging. One of the main mechanisms of aging regulation in budding yeast is replicative aging, in which the mother cell ages with every budding event, by asymmetrically sequestering damaged proteins, thereby “resetting” the aging clock of the newly-emerged daughter cell. Although a number of tools already exist to both track individual yeast cells over time and to isolate successive generations of yeast, these tools are costly, time-consuming, and not easily scalable. Sorting based on fluidic forces confers the advantages of system simplicity, high-throughput work and low cost. Replicative age dependent separation of yeast requires characterization of the physical and biological properties of the cells, not previously done. An additional challenge is the size variance in each age group, followed by size overlap of different age groups. To withstand the challenge of replicative age dependent sorting of yeast, we coupled the unique phenomena of cross-section preferential concentration by Dean vortices with opposing shear induced lift forces, both occur in extremely high Reynolds numbers. We focused different size yeast to distant locations, to achieve biologically significant age dependent separation of the yeast despite the considerable age variance within each size binned group. Separation efficiency was monitored by yeast image processing, and replicative age dependent protein degradation was measured and analyzed.