Vincent Studer, P-O Strale et al. dans Advanced Materials

Multiprotein Printing by Light-Induced Molecular Adsorption. Strale PO, Azioune A, Bugnicourt G, Lecomte Y, Chahid M, Studer V. Adv Mater . 2015 Dec 21. doi: 10.1002/adma.201504154. [Epub ahead of print] 

Vincent Studer:  Fast, affordable and reproducible cell patterning technologies are required for the future development of high-throughput cells-based assays and tissue engineering.  In our paper published in Advanced Materials, we describe how to perform high resolution, multi-adhesion protein micro-patterning using a new technology named “Light-Induced Molecular Adsorption” (LIMA) allowing an exquisite control over different cell types populations positioning. Our LIMA method is based on a water-soluble photo-initiator which is able to reverse the antifouling property of polymer brushes when exposed to UV light.

We showed that the density of adsorbed molecules scales almost linearly with the dose of UV light. LIMA is also well adapted to wide field illumination schemes, consequently micron-scale patterns where adhesion proteins can adsorb, are printed within seconds. Moreover, the very low background of these patterns allows for the sequential printing of multiple proteins. Our optical set-up (widefield DMD-based projection system coupled to a conventional epifluorescence microscope) allows us to generate arbitrary grayscale patterns of UV light and thus controlled gradients of multiple biomolecules. As a demonstration of the robustness of LIMA protein printing, we reproduced “the birth of Venus” of Botticelli by superimposing complex patterned of three different fluorescent molecules (see below , zoomable picture).

A: Principle of the LIMA method allowing for orthogonal patterning of two different fluorescent proteins as shown in B. Scale Bar = 50 µm. C: Epifluorescence image of a eproduction “of the Birth of Venus” by sequential of printing of three different fluorescent molecules. Scale Bar = 100µm.

We showed that the range of application of our LIMA system extends from the single molecule up to the multicellular scale: micropatterns of individual fluorescent molecules provide a tool for the quantitative study of biomolecular interactions. At larger scale, the ability to rapidly generate complex and dynamic protein landscapes enable studies of single cell dynamics, and of cell-cell and cell-matrix interaction.

Altogether, light-induced molecular adsorption of adhesion proteins provides a generic method for fast, high resolution patterning of multiple proteins allowing unmatched orthogonal cell printing capabilities, which turn LIMA into a very powerful method for multi-scale co-culture with straightforward applications in tissue engineering and biomedical research.