Absolute 3D reconstruction of thin films topography in microfluidic channels by interference reflection microscopy.

The travel of droplets, bubbles, vesicles, capsules, living cells or small organisms in microchannels is a hallmark in microfluidics applications. A full description of the dynamics of such objects requires a quantitative understanding of the complex hydrodynamic and interfacial interactions between objects and channel walls. In this paper, we present an interferometric method that allows absolute topographic reconstruction of the interspace between an object and channel walls for objects confined in microfluidic channels. Wide field microscopic imaging in reflection interference contrast mode (RICM) is directly performed at the bottom wall of microfluidic chips. Importantly, we show that the reflections at both the lower and upper surface of the microchannel have to be considered in the quantitative analysis of the optical signal. More precisely, the contribution of the reflection at the upper surface is weighted depending on the light coherence length and channel height. Using several wavelengths and illumination apertures, our method allows reconstructing the topography of thin films on channel walls in a range of 0-500 nm, with a precision as accurate as 2 nm for the thinnest films. A complete description of the protocol is exemplified for oil in water droplets travelling in channels of height 10-400 µm at a speed up to 5 mm s(-1).

New modeling of reflection interference contrast microscopy including polarization and numerical aperture effects: application to nanometric distance measurements and object profile reconstruction.

We have developed a new and improved optical model of reflection interference contrast microscopy (RICM) to determine with a precision of a few nanometers the absolute thickness h of thin films on a flat surface in immersed conditions. The model takes into account multiple reflections between a planar surface and a multistratified object, finite aperture illumination (INA), and, for the first time, the polarization of light. RICM intensity I is typically oscillating with h. We introduce a new normalization procedure that uses the intensity extrema of the same oscillation order for both experimental and theoretical intensity values and permits us to avoid significant error in the absolute height determination, especially at high INA. We also show how the problem of solution degeneracy can be solved by taking pictures at two different INA values. The model is applied to filled polystyrene beads and giant unilamellar vesicles of radius 10-40 microm sitting on a glass substrate. The RICM profiles I(h) can be fitted for up to two to three oscillation orders, and extrema positions are correct for up to five to seven oscillation orders. The precision of the absolute distance and of the shape of objects near a substrate is about 5 nm in a range from 0 to 500 nm, even under large numerical aperture conditions. The method is especially valuable for dynamic RICM experiments and with living cells where large illumination apertures are required.

Surface enhanced ellipsometric contrast (SEEC) basic theory and lambda/4 multilayered solutions.

The fundamentals of a new high contrast technique for optical microscopy, named "Surface Enhanced Ellipsometric Contrast" (SEEC), are presented. The technique is based on the association of enhancing contrast surfaces as sample stages and microscope observation between cross polarizers. The surfaces are designed to become anti-reflecting when used in these conditions. They are defined by the simple equation r(p) + r(s) = 0 between their two Fresnel coefficients. Most often, this equation can be met by covering a solid surface with a single lambda/4 layer with a well defined refractive index. A higher flexibility is obtained with multilayer stacks. Solutions with an arbitrary number of all-dielectric lambda/4 layers are derived.

Stability of thin nematic films. Commentary on "Morphology and structure of thin liquid-crystalline films at nematic-isotropic transition" by P. Ziherl and S. Zumer.

We discuss the peculiarity of thin nematic films on solid substrates with a free surface, underlining the differences with what is usually seen in dewetting. We review the thermodynamic basis of the coupled phase/thickness separation that has previously been shown experimentally. We give new experimental evidences for the origin of the coupling force chosen in our previous theoretical model. This additional information contributes to the discussion raised by the article of Ziherl and Zumer in this issue.

Binary separation in very thin nematic films: thickness and phase coexistence.

The behavior as a function of temperature of very thin films (10 to 200 nm) of pentylcyanobiphenyl on silicon substrates is reported. In the vicinity of the nematic-isotropic transition we observe a coexistence of two regions of different thicknesses: thick regions are in the nematic state while thin ones are in the isotropic state. Moreover, the transition temperature is shifted downward following a 1/h(2) law ( h is the film thickness). Microscope observations and small-angle x-ray scattering allowed us to draw a phase diagram which is explained in terms of a binary first-order phase transition where thickness plays the role of an order parameter.

Organization of cyanobiphenyl liquid crystal molecules in prewetting films spreading on silicon wafers.

We observe prewetting films of 8CB (4'-n-octyl-4-cyanobiphenyl) spreading at room temperature on silicon wafers by ellipsometry and x-ray reflectivity. Ellipsometry indicates the formation of a nondense monolayer spreading in front of a 45-A-thick film. X-ray reflectivity, performed using a ribbon geometry for the liquid crystal (LC) reservoir, allows us to determine the organization of the 8CB molecules in the homogenous film. It consists of a trilayer stacking with a smecticlike bilayer standing above a polar monolayer with tilted molecules. We show that the thickness of the bilayer is equal to the smectic periodicity in the bulk material and that the tilt angle of the molecules in contact with the solid surface is close to 60 degrees, in good agreement with second-harmonic generation studies reported by other groups. Such organization can be precisely determined using x-ray reflectivity because it induces a modulation of the electron density along the normal to the surface. Furthermore, a study of the ellispometric profile of a drop heated in the nematic phase, where we observe a complete spreading of the LC, shows the complex structuration of the LC close to the solid interface. In particular, the spreading behavior of the trilayer compared to the subsequent smecticlike bilayers indicates the existence of specific interaction between the trilayer and silicon wafer.