Improvement of yeast biochip sensitivity using multilayer inorganic sol-gel substrates.

Today, microarray fluorescence detection is still limited because a great proportion of hybrids remain undetectable. In this paper we describe sol-gel optical multilayers (stacks of low- and high-index layers) deposited on glass slides which increase the fluorescence of DNA microarrays and favour the detection of fluorescent targets. An alternative to the expensive and time-consuming physical vapour deposition technology is proposed. It is a low-cost sol-gel coating of glass slides, each layer being made by "dipping" (alternatively in SiO2 or TiO2 solutions), "draining and drying". After the selection of the best surface layer of the substrates, the multilayer mirrors modelled for one (Cy3) or two (Cy3 and Cy5) fluorophores are spotted with a series of Yeast probes and compared to similar microarrays on standard glass slides through hybridisation experiments. The fluorescence images of the mirrors show increased signals for all the probes. The enhancement factors determined for Cy3 and for Cy3/Cy5 mirrors (10-12 and 4-5, respectively) are consistent with the initial modelling. This allows the assessment of the basal expression levels of Yeast low-expressed genes. Moreover, these substrates show a noticeable increase in sensitivity for induction/repression ratio measurements in differential gene expression experiments. So, they could be considered as promising tools for the analysis of small biological samples.

Polarization and particle size dependence of radiative forces on small metallic particles in evanescent optical fields. Evidences for either repulsive or attractive gradient forces.

We have observed the motion of metallic particles above various optical waveguides injected by 1064nm radiation. Small gold particles (250nm diameter) are attracted towards the waveguide where the intensity of the optical field is maximum, and are propelled at high velocity (up to 350mum/s) along the waveguide due to radiation pressure. The behaviour of larger metallic particles (diameter >600nm) depends on the polarization of the evanescent field: for TM polarization they are attracted above the waveguide and propelled by the radiation pressure; for TE polarization they are expelled on the side of the waveguide and propelled at much smaller velocity. This is consistent with calculations of radiative forces on metallic particles by Nieto-Vesperinas et al. 3D-finite element method calculations carried out for our experimental situations confirm the observed dependence with the polarization of the field and the size of the particles. These observations open the way to the development of new microsystems for particles manipulations and sorting applications.

Competitive technology approaches for electronic hybridization detection in a microsystem with microfluidics for diagnosis genetic tests.

This paper is presenting competitive technology alternatives for the electronic hybridization detection in a microsystem with microfluidics for diagnosis genetic tests that are carried out by two competitive research projects. The technologies developed are a photosensor, a capacitive sensor and an optical real-time affinity biosensor. The performance of those biosensors will be evaluated but also their manufacturability and cost will define the appropriateness of each one for industrialization and their integration on a microsystem for diagnosis genetic testing.

Multiple wavelength fluorescence enhancement on glass substrates for biochip and cell analyses.

In microarrays experiments, a serious limitation is the unreliability of low signal intensities data and the lack of reproducibility for the resulting ratios between samples and controls. Most of the light emitted by a fluorophore at the air/glass interface of a glass slide is absorbed by the glass so just a part of the emitted fluorescence is detected. To improve the sensitivity of the fluorescence detection of both common fluorophores Cy3 and Cy5 in DNA microarrays and fluorescent cell analyses, we have designed a multi layer mirror with alternative thin layers of SiO2 and HfO2. This mirror (MOTL) prevents fluorescence absorption, allows the simultaneous enhancement of the fluorescence signals and increases the dynamic range of the slides. Using MOTL slides, Cy3 and Cy5 intensities are enhanced by 5-8-fold, consequently, the fluorescence analysis becomes easier and should allow the detection of low copy number genes or weakly fluorescent cells. With the same approach, other multiple optical thin layer slides could be designed for other series of fluorophores, extending the field of their applications.

Optical manipulation of microparticles and cells on silicon nitride waveguides.

We demonstrate the optical manipulation of cells and dielectric particles on the surface of silicon nitride waveguides. Glass particles with 2microm diameter are propelled at velocities of 15microm/s with a guided power of 20mW. This is approximately 20 times more efficient than previously reported, and permits to use this device on low refractive index objects such as cells. Red blood cells and yeast cells can be trapped on the waveguide and pushed along it by the action of optical forces. This kind of system can easily be combined with various integrated optical structures and opens the way to the development of new microsystems for cell sorting applications.