Multiwell plate tools for controlling cellular alignment with grooved topography.

In many tissues, cells must be aligned for proper function. This alignment can occur at the cellular and/or subcellular (protein/molecular) level. The alignment of cytoskeletal components, in fact, precedes whole cell alignment. A variety of methods exist to manipulate cytoskeletal and whole cell alignment; one of the simplest and most predictable involves seeding adherent cells onto defined substrate topography. We present here two methods to create grooved multiwell plates: one involving microfabrication, which allows for custom design of substrate topography, and a simpler, inexpensive method using commercially available diffraction gratings. We also include methods for manual and automatic quantification of cell alignment.