Computer simulations of the energy dissipation rate in a fluorescence-activated cell sorter: Implications to cells |
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Authors: | Mollet Mike Godoy-Silva Ruben Berdugo Claudia Chalmers Jeffrey J |
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Affiliation: | Department of Chemical and Biomolecular Engineering, The Ohio State University, 125 Koffolt Laboratories, 140 W 19th Ave, Columbus, Ohio 43210, USA. |
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Abstract: | Fluorescence activated cell sorting, FACS, is a widely used method to sort subpopulations of cells to high purities. To achieve relatively high sorting speeds, FACS instruments operate by forcing suspended cells to flow in a single file line through a laser(s) beam(s). Subsequently, this flow stream breaks up into individual drops which can be charged and deflected into multiple collection streams. Previous work by Ma et al. (2002) and Mollet et al. (2007; Biotechnol Bioeng 98:772-788) indicates that subjecting cells to hydrodynamic forces consisting of both high extensional and shear components in micro-channels results in significant cell damage. Using the fluid dynamics software FLUENT, computer simulations of typical fluid flow through the nozzle of a BD FACSVantage indicate that hydrodynamic forces, quantified using the scalar parameter energy dissipation rate, are similar in the FACS nozzle to levels reported to create significant cell damage in micro-channels. Experimental studies in the FACSVantage, operated under the same conditions as the simulations confirmed significant cell damage in two cell lines, Chinese Hamster Ovary cells (CHO) and THP1, a human acute monocytic leukemia cell line. |
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Keywords: | FACS flow cytometry EDR cell damage shear stress CHO |
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