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 patterns.
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http://dx.doi.org/10.1021/la2039202 | DOI Listing |
ACS Appl Mater Interfaces
January 2018
The School of Physics, University of Sydney, Sydney, New South Wales 2006, Australia.
Multiprotein micropatterning allows the creation of complex, controlled microenvironments for single cells that can be used for the study of the localized effects of various proteins and signals on cell survival, development, and functions. To enable analysis of cell interactions with microprinted proteins, the multiprotein micropattern must have low cross-contamination and high long-term stability in a cell culture medium. To achieve this, we employed an optimized plasma ion immersion implantation (PIII) treatment to provide polystyrene (PS) with the ability to covalently immobilize proteins on contact while retaining sufficient transparency and suitable surface properties for contact printing and retention of protein activity.
View Article and Find Full Text PDFPLoS One
February 2015
Department of Biochemistry, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands.
T cell signaling is triggered through stimulation of the T cell receptor and costimulatory receptors. Receptor activation leads to the formation of membrane-proximal protein microclusters. These clusters undergo tyrosine phosphorylation and organize multiprotein complexes thereby acting as molecular signaling platforms.
View Article and Find Full Text PDFSmall
October 2013
Institute of Nanotechnology (INT) and Karlsruhe, Nano Micro Facility (KNMF), Karlsruhe Institute of Technology (KIT), Germany; Physical Institute and Center for Nanotechnology (CeNTech), University of Münster, Germany.
Multiplexing, i.e., the application and integration of more than one ink in an interdigitated microscale pattern, is still a challenge for microcontact printing (μCP) and similar techniques.
View Article and Find Full Text PDFLangmuir
January 2012
Department of Bioengineering, University of Utah, Salt Lake City, Utah 84112, USA.
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.
View Article and Find Full Text PDFAnal Chem
June 2006
Division of Molecular and Applied Physics, Department of Physics, Chemistry, and Biology, Linköping University, S-581 83 Linköping, Sweden.
The fabrication of a novel biochip, designed for dissection of multiprotein complex formation, is reported. An array of metal chelators has been produced by piezo dispensing of a bis-nitrilotriacetic acid (bis-NTA) thiol on evaporated gold thin films, prestructured with a microcontact printed grid of eicosanethiols. The bis-NTA thiol is mixed in various proportions with an inert, tri(ethylene glycol) hexadecane thiol, and the thickness and morphological homogeneity of the dispensed layers are characterized by imaging ellipsometry before and after back-filling with the same inert thiol and subsequent rinsing.
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