Franck Thibaudau

Cell adhesion is an important cellular process and is mediated by adhesion proteins residing on the cell membrane. Sometimes, two types of linker proteins are involved in adhesion, and they can segregate to form domains through a poorly understood size-exclusion process. We present an experimental and theoretical study of adhesion via linkers of two different sizes, realised in a mimetic model-system, based on giant unilamellar vesicles interacting with supported lipid bilayers. Here, adhesion is mediated by DNA linkers with two different lengths, but with the same binding enthalpy. We study the organisation of these linkers into domains as a function of relative fraction of long and short DNA constructs. Experimentally, we find that, irrespective of the composition, the adhesion domains are uniform with coexisting DNA bridge types, despite their relative difference in length of 9 nm. However, simulations suggest formation of nanodomains of the minority fraction at short length scales, which is below the optical resolution of the microscope. The nano-aggregation is more significant for long bridges, which are also more stable.

Adhesion domains forming at the membrane interfaces between two cells or a cell and the ex-tracellular matrix commonly involve multiple proteins bridges. However, the physical mechanisms governing the domain structures are not yet fully resolved. Here we present a joint experimental and theoretical study of a mimetic model-system, based on giant unilammelar vesicles interacting with supported lipid bilayers, with which the underlying physical effects can be clearly identified. In our case, adhesion is induced by simultaneous action of DNA linkers with two different lengths. We study the organization of bridges into domains as a function of relative fraction of long and short DNA constructs. Irrespective of the composition, we systematically find adhesion domains with coexisting DNA bridge types, despite their relative differences in length of 9 nm. However, at short length scales, below the optical resolution of the microscope, simulations suggest the formation of nanodomains by the minority fraction. The nano-aggregation is more significant for long bridges, which are also more stable, even though the enthalpy of membrane insertion is the same for both species.

We report results on ultrafast photothermal release of DNA from gold nanoparticles. We show that dehybridization of oligonucleotide duplex anchored on a gold nanoparticle surface occurs during a single laser pulse, leading to the release of single-strand DNA in solution. Breaking of the Au–S bond anchoring the duplex and the release of thiolated DNA are also evidenced. Our findings show that the size distribution of the nanoparticles plays a major role in the control of both phenomena. We establish a criterion regarding the size distribution of nanoparticles that allows full release of DNA without breaking of the anchoring thiol bonds.

Aminosilane film on mica is investigated in this work for further use as a substrate for DNA immobilization. We study the deposition of N-(2-aminoethyl)-3-aminopropyltrimethoxysilane (AAPS) on mica surfaces by atomic force microscopy, X-ray photoelectron spectroscopy, and contact angle measurements. Deposition both in aqueous solution and in vapor-phase is explored. In aqueous solution, it is found that the interactions of molecules with the mica surface are pH-dependent. In the case of vapor phase deposition, a multilayer film composed of a first monolayer and successive bilayers is obtained. This is the first observation of the organization of aminosilane films on mica.

A label-free two-dimensional colorimetric DNA sensor is reported. This sensor is based on the 2D aggregation of oligonucleotide-modified gold nanoparticle probes induced by the molecular hybridization of single-stranded oligonucleotide probes and their complementary single-stranded DNA targets. To detect the aggregation, we have developed a new detection method based on the selective desorption of nonaggregated nanoparticles. We will show here that this detection method is highly specific and allows the quantification of the DNA targets.