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Microfluidic phenotyping methods have been of vital importance for cellular characterization, especially for evaluating single cells. In order to study the deformability of a single cell, we devised and tested a tunable microfluidic chip-based method. A pneumatic polymer polydimethylsiloxane (PDMS) membrane was designed and fabricated abutting a single-cell trapping structure, so the cell could be squeezed controllably in a lateral direction. Cell contour changes under increasing pressure were recorded, enabling the deformation degree of different types of single cell to be analyzed and compared using computer vision. This provides a new perspective for studying mechanical properties of cells at the single cell level.