Dual pipette aspiration: a unique tool for studying intercellular adhesion.

The dual pipette aspiration (DPA) assay is a highly versatile tool that enables the micromanipulation of cells and the precise measurement of a range of biophysical parameters in combination with concurrent high-resolution imaging. DPA permits the juxtaposition of cells, their manipulation using pressure and the controlled formation or separation of cell-cell contacts. The DPA set-up can thus readily be used to probe the dynamics and mechanics of cell-cell adhesion, notably adhesion strength and adhesion energy. In particular, the DPA set-up has been used to measure a wide range of separation forces between pairs of cells. Here, we describe how to build and use the DPA set-up in order to measure the separation force of cell doublets. We first describe how to prepare adequate pipettes, then how to assemble and calibrate the pipettes and pressure control devices, followed by how to manipulate cells in order to calculate separation forces. Finally, we give recommendations on how to use the DPA set-up and compare it to other methods used to study cell-cell contacts and adhesion strength in particular.