Figure 5. Microfluidic dissociation augments trypsin and EDTA treatments.

Total recovery (A-C) and single cell percentage (D-F) for different spheroid types following treatment with trypsin for 5 min, EDTA for 5 min, or EDTA for 15 min, with or without microfluidic device processing at 12.5 mL/min. Controls received mechanical treatment by pipetting and vortexing. (A,D) For HCT 116 spheroids, the device increased total recovery for both treatments by two- to three-fold, with improved single cell content as well. Maximal results were obtained after 1 pass for the trypsin case, while additional passes enhanced EDTA results. (B,E) For NCI-H1650 spheroids, samples were only partially dissociated for brief trypsin and EDTA treatments, but 10 passes through the microfluidic device improved total recovery by more than two-fold. (C,F) For LS 174T spheroids, brief trypsin and EDTA treatments were extremely inefficient, but were enhanced by more than 10-fold following microfluidic device processing (10 passes). All total count results were normalized to the maximum count determined by fully digesting each spheroid type with trypsin (see Supporting Information). Error represent the standard error from at least three independent experiments.