HeLa cells (CCL-2, American Type Culture Collection, USA) were cultured using cell culture media (Eagle’s minimum essential medium modified with 10% fetal bovine serum) in a 37°C cell culture incubator. Before the experiment, the cultured cells were suspended in the same culture media and transferred to the acoustic fluidic chamber with a pipette. The experiments were conducted immediately after the transfer to prevent adhesion of cells to the substrate. Silica beads (Sigma-Aldrich, USA) and polystyrene beads (Bangs Laboratories Inc., USA) were diluted with deionized water to appropriate concentrations for particle patterning and streaming tracking purposes.

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