This collection comprises raw data and media files supporting the PNAS manuscript, "Robust acoustic trapping and perturbation of single-cell microswimmers illuminate three-dimensional swimming and ciliary coordination." The unicellular alga Chlamydomonas reinhardtii is widely used to investigate the mechanisms underlying ciliary beating and coordination. Freely swimming cells are difficult to image with sufficient resolution to capture cilia motion, necessitating that the cell body be held during experiments. Acoustic confinement enables investigation of cell and cilia motion under conditions that approximate free swimming. The video files in this repository entry are used to measure cell and cilia behavior under acoustically trapped and free swimming conditions.