Figure 5.

At the base of each macrowell on a 96-well plate are ~300 microwells in a grid. Pictured here are microwells that are ~40 μm in diameter and spaced 240 μm apart. Cells are loaded into each macrowell, and they settle into the microwells over time. Loading efficiency can be checked using a microscope by visualizing a grid of loaded microwells among the off-grid/excess cells (examples of loaded wells are marked by the blue arrows). Following the wash step, excess cells are removed, leaving behind an array of patterned cells. For these images, 40,000 TK6 cells were added to each macrowell in a volume of 80 μL. Cells remain trapped in wells following 20 min of loading time. Note that loading time can be optimized for different cell types, but can be as low as 5 min.