USE OF A SEPARABILITY PARAMETER FOR THE DESIGN OF A HIGH FREQUENCY DIELECTROPHORESIS CELL SORTER DEVICE

摘要

We present a method to quantify and enhance separation of binary cells mixture in the microfluidic device using high frequency dielectrophoresis (＞20 MHz). At these frequencies, the DEP response depends primarily on the dielectric properties of the cytoplasm. In order to achieve efficient separation, there must be a difference in the intrinsic dielectric properties of the populations to be sorted. For algae cells, the shift in high frequency dielectrophoresis response during lipid accumulation can be used as a basis of separation. We defined a separability parameter based on the expected difference in the dielectrophoresis responses of the algae cells. Chlamydomonas reinhardtii cells were cultured in regular media and then the same cells were cultured under nitrogen-free conditions to accumulate neutral (non-polar) lipids. Separability of microalgae cells with different lipid content via high frequency dielectrophoresis were investigated by a thin needle shaped electrodes patterned by standard photolithographic and wet etching procedures. Experimental separability factors were measured by estimation of relative lipid content with BODIPY 505/515 fluorescence dye and calculating the area-weighted intensity average of fluorescent images. Theoretical separability parameter was calculated using analytical analysis of single shell model by MATLAB. Theoretical and experimental separability parameters, as tools to determine the optimal separation method, were calculated for microalgae cells with different lipid content. This objective function was maximized in the range of 35-45 MHz for C. reinhardtii cells after 21 days of lipid accumulation in a static separation. In order to design a continuous cell sorter device, the theoretical separation factor was maximized based on differences in the magnitude or the direction of the DEP force.