| Summary: | A cell filtration platform that affords accurate size separation and minimizes fouling was developed. The platform features an ultra-thin porous membrane (UTM) filter, a pumping head filtration with backflush (PHF), and cell size measurement (CSM) software. The UTM chip is an ultrathin free-standing membrane with a large window area of 0.68 mm<sup>2</sup>, a pore diameter of 5 to 9 μm, and a thickness of less than 0.9 μm. The PHF prevents filter fouling. The CSM software analyzes the size distributions of the supernatants and subnatants of isolated cells and presents the data visually. The <i>D</i>99 particle size of cells of the chronic myeloid leukemia (CML) line K562 decreased from 22.2 to 17.5 μm after passage through a 5-μm filter. K562 cells could be separated by careful selection of the pore size; the recovery rate attained 91.3%. The method was compared to conventional blocking models by evaluating the mean square errors (MSEs) between the measured and calculated filtering volumes. The filtering rate was fitted by a linear regression model with a significance that exceeded 0.99 based on the <i>R</i><sup>2</sup> value. The platform can be used to separate various soft biomaterials and afford excellent stability during filtration.
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