|One of our group's main topics is the generation of bioactive surfaces for application in cell culture experiments. The structured immobilization of biological active compounds (proteins as well as small molecules) on surfaces in length scales smaller than a cell enables many interesting applications in biological and biomedical research. Because of the mild process parameters and the ability of writing different inks within one pattern ("multiplexing"), DPN offers ideal possibilities for the fabrication of complex bioactive patterns. DPN with phospholipids (lipid DPN) enables the parallel immobiliation of very sensitive proteins by introducing specific binding sites (here over nickel-chelating head groups for binding of his-tag-proteins and biotinyliated headgroups for building up a streptavidin sandwich, scheme shown below). An example for the presentation of small bioactive molecules is given by our research on mast cells. Patterns with different ammount of admixing of a model allergen (dinitrophenol) are written by lipid DPN to study the activation of mast cells as well as the suppression of the response by glucocorticoids (upper right). We also employ polymer pen lithography (PPL) to print bioactive substances and proteins (bottom right). PPL combines advantages of DPN (multiplexing, high-precision, mild process parameters) with those of microcontact printing (inexpensive stamps, large area) making it a great tool for generation of multi component bioactive surface patterns.||Mast cells on an allergenic DPN structure (left) and on a non-activating control pattern (right). The green fluorescence shows the co-localization of the allergen-recognizing cell receptor with the allergenic pattern.
Schematic representation of the multi-color PPL printing process. The inset shows an actual fluorescence image of a pattern generated with three different fluorescently labeled phospholipids.
Scheme of the binding strategy for multiplexed protein deposition via self-assembly on lipid DPN generated specific binding sites. The insets show fluorescence microscopy images of the realization of this concept with his-GFP (green) and Cy3-streptavidin (red).
Interdigitated Multicolored Bioink Micropatterns by Multiplexed Polymer Pen Lithography
F. Brinkmann, M. Hirtz, A. M. Greiner, M. Weschenfelder, B. Waterkotte, M. Bastmeyer, H. Fuchs
Small 9 (2013) 3266-3275, DOI:10.1002/smll.201203183
Allergen arrays for antibody screening and immune cell activation profiling generated by parallel lipid dip-pen nanolithography
S. Sekula-Neuner, J. Maier, E. Oppong, E.; A. C. B. Cato, M. Hirtz, H. Fuchs
Small 8 (2012) 585-591, DOI:10.1002/smll.201101694
Multiplexed lipid dip-pen nanolithography on subcellular scales for the templating of functional proteins and cell culture
S. Sekula, J. Fuchs, S. Weg-Remers, P. Nagel, S. Schuppler, J. Fragala, N. Theilacker, M. Franzreb, C. Wingren, P. Ellmark, C. A. K. Borrebaeck, C. A. Mirkin, H. Fuchs, S. Lenhert
Small 4 (2008) 1785-1793, DOI:10.1002/smll.200800949