  Micropatterning techniques have become an important tool for the study of
cell behavior in controlled microenvironments. As a consequence, several
approaches for the creation of micropatterns have been developed in recent
years. However, the diversity of substrates, coatings and complex patterns used
in cell science is so great that no single existing technique is capable of
fabricating designs suitable for all experimental conditions. Hence, there is a
need for patterning protocols that are flexible with regard to the materials
used and compatible with different patterning strategies to create more
elaborate setups. In this work, we present a versatile approach to
micropatterning. The protocol is based on plasma treatment, protein coating,
and a PLL-PEG backfill step, and produces homogeneous patterns on a variety of
substrates. Protein density within the patterns can be controlled, and density
gradients of surface-bound protein can be formed. Moreover, by combining the
method with microcontact printing, it is possible to generate patterns composed
of three different components within one iteration of the protocol. The
technique is simple to implement and should enable cell science labs to create
a broad range of complex and highly specialized microenvironments.
