Separation of viable and non-viable yeast using dielectrophoresis

Abstract

Dielectrophoresis, the movement of particles in non-uniform AC electric fields, was used to rapidly separate viable and non-viable yeast cells with good efficiency. Known mixtures of viable and heat-treated cells of Saccharomyces cerevisiae were separated and selectively isolated using positive and negative dielectrophoretic forces generated by microelectrodes in a small chamber. Good correlations with the initial known relative compositions were obtained by direct microscopic counting of cells at the electrodes after initial dielectrophoretic separation ( r = 0.995), from methylene blue staining ( r = 0.992) and by optical absorption measurements ( r = 0.980) of the effluent after selectively flushing out the viable and non-viable cells from the chamber. Through measurement of cell viability by staining with methylene blue and plate counts, for an initial suspension of approx. 1.4 × 10 7 cells per ml containing 60% non-viable cells, the dielectrophoretically separated non-viable fraction contained 3% viable cells and the viable fraction 8% dead cells. The separation efficiency is increased by dilution of the initial suspension or by repeat operation(s). Cell viability was not affected by the separation procedure.