High Fidelity Nanopatterning of Proteins onto Well-Defined Surfaces Through Subtractive Contact Printing

Abstract

In the pursuit to develop enhanced technologies for cellular bioassays as well as understand single cell interactions with its underlying substrate, the field of biotechnology has extensively utilized lithographic techniques to spatially pattern proteins onto surfaces in user-defined geometries. Microcontact printing (μCP) remains an incredibly useful patterning method due to its inexpensive nature, scalability, and the lack of considerable use of specialized clean room equipment. However, as new technologies emerge that necessitate various nano-sized areas of deposited proteins, traditional microcontact printing methods may not be able to supply users with the needed resolution size. Recently, our group developed a modified “subtractive microcontact printing” method which still retains many of the benefits offered by conventional μCP. Using this technique, we have been able to reach resolution sizes of fibronectin as small as 250 nm in largely spaced arrays for cell culture. In this communication, we present a detailed description of our subtractive μCP procedure that expands on many of the little tips and tricks that together make this procedure an easy and effective method for controlling protein patterning.

Multifunctional Structured Platforms: From Patterning of Polymer-Based Films to Their Subsequent Filling with Various Nanomaterials

“…Microcontact printing (μCP) is a soft lithographic method that creates micrometer (or nanometer, e.g., nanocontact printing) surface relief patterns onto well-defined polymeric surfaces through the conformal contact of the latter with various patterns, of a PDMS master stamp, that are covered with polymer-based or self-assembled monolayer (SAM) inks ( Figure 7 a). Tips and tricks on how to explore the maximum potential of the μCP technique can be found in the literature [ 142 ]. This patterning technique is preponderantly used in biological applications, and it is accompanied by processes such as functionalization [ 143 ].…”

Polymeric and biological membranes for organ-on-a-chip devices

“…The selective transfer is achieved via microcontact printing (μCP) or microfluidics and provides geometrical cues to which cells (e.g., cardiac tissue or muscular fibers) respond through changes in direction and alignment 124 . μCP protein patterns can reach sizes as low as 0.5 μm; however, pattern integrity depends on the feature size and the chemical nature of the membrane, as larger features and hydrophobic surfaces present more robust binding 125 , 126 . A study conducted by Wright and collaborators achieved fibronectin patterning onto PDMS and polystyrene with reusable 10-μm-thick parylene-C stencils (Fig.…”