Depositing multiple proteins on the same substrate in positions similar
to the natural cellular environment is essential to tissue engineering and regenerative
medicine. In this study, the development and verification of a multiprotein
microcontact printing (μCP) technique is described. It is shown that patterns of
multiple proteins can be created by the sequential printing of proteins with
micrometer precision in registration using an inverted microscope. Soft polymeric
stamps were fabricated and mounted on a microscope stage while the substrate to be
stamped was placed on a microscope objective and kept at its focal distance. This
geometry allowed for visualization of patterns during the multiple stamping events and
facilitated the alignment of multiple stamped patterns. Astrocytes were cultured over
stamped lane patterns and were seen to interact and align with the underlying protein pattern
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