Abstract: The isolation of single cells is challenging and an essential step in many biomedical workflows, for example genetic analysis and drug-based assays. A common method for isolating single cells is through limiting dilution; however, dark regions at the periphery of standard microtiter wells make it difficult for users to be sure a well contains just one cell. Researchers often perform a second round of limiting dilution to increase confidence in monoclonality, adding time and cost to experiments. An elegant solution is to add cells to sessile drops with low contact angles. The minimal refraction of light across the drop-air interface allows users to obtain a clear image of a single cell within the drop using a standard microscope. Such drops are deposited in the base of microtiter wells and cell presence is confirmed microscopically. This allows users to be certain a well contains a single cell. We present a method for automated generation of shallow drops and cell deposition in standard 96-well plates. We confirm the presence of a single cell per well (using human embryonic kidney, HEK, and prostate cancer, LNCaP, cells), and that cells subsequently grow normally after imaging. This also provides a reliable measure of cloning efficiency without being subject to uncertainties introduced by Poisson statistics. In addition, colonies are more likely to grow in the centre of a well when compared to those generated by standard limiting dilution, and images are not compromised by edge effects at the well periphery. We also show a method to pick a single cell from such drops, allowing biologists to choose which cells to use in subsequent assays. We add ~100 HEK cells to a drop and withdraw volume to determine picking efficiency. Reliable picking is achieved by keeping cells close to the drop centre. By adjusting drop volume, we reduce cell dispersion and obtain picking efficiencies > 95%. This demonstrates that sessile drops are a promising alternative to expensive and time-consuming methods for single-cell isolation. Temporarily holding a cell in a shallow drop allows users to identify a single cell, and then pick the cell with confidence for downstream cloning or assay.