Abstract
Immune cells continuously adapt their mechanical properties for proper circulation and elicitation of immune responses. Here, we provide a step-by-step protocol for probing the single-cell mechanical properties of primary human monocytes using acoustic force spectroscopy (AFS). We describe steps for the calibration of the AFS chips, the isolation of monocytes from buffy coats, and the probing of monocyte mechanics using AFS. We then detail the data analysis strategy. The protocol is useful for characterizing a wide range of immune cells under various conditions in physiology and pathology. For complete details on the use and execution of this protocol, please refer to Evers et al. 1 and Evers et al.2.