Thomas Dehoux

Picosecond time scale imaging of mechanical contacts

By means of an ultrafast opto-acoustic technique we study the nanoindentation of thin chromium films on sapphire substrates using a ceramic ball bearing. Acoustic pulses at approximately 40 GHz returning from the film-indenter interface allow the film indentation profiles to be probed to sub-nanometer resolution over contact areas approximately 25 microm in radius. The deformation of the films during loading is thereby revealed. Furthermore, thermal wave imaging of the contact at megahertz frequencies is simultaneously achieved.

Transverse mechanical properties of cell walls of single living plant cells probed by laser-generated acoustic waves.

Probing the mechanical properties of plant cell wall is crucial to understand tissue dynamics. However, the exact symmetry of the mechanical properties of this anisotropic fiber-reinforced composite remains uncertain. For this reason, biologically relevant measurements of the stiffness coefficients on individual living cells are a challenge. For this purpose, we have developed the single-cell optoacoustic nanoprobe (SCOPE) technique, which uses laser-generated acoustic waves to probe the stiffness, thickness and viscosity of live single-cell subcompartments. This all-optical technique offers a sub-micrometer lateral resolution, nanometer in-depth resolution, and allows the non-contact measurement of the mechanical properties of live turgid tissues without any assumption of mechanical symmetry. SCOPE experiments reveal that single-cell wall transverse stiffness in the direction perpendicular to the epidermis layer of onion cells is close to that of cellulose. This observation demonstrates that cellulose microfibrils are the main load-bearing structure in this direction, and suggests strong bonding of microfibrils by hemicelluloses. Altogether our measurement of the viscosity at high frequencies suggests that the rheology of the wall is dominated by glass-like dynamics. From a comparison with literature, we attribute this behavior to the influence of the pectin matrix. SCOPE’s ability to unravel cell rheology and cell anisotropy defines a new class of experiments to enlighten cell nano-mechanics.

All-optical broadband ultrasonography of single cells.

Cell mechanics play a key role in several fundamental biological processes, such as migration, proliferation, differentiation and tissue morphogenesis. In addition, many diseased conditions of the cell are correlated with altered cell mechanics, as in the case of cancer progression. For this there is much interest in methods that can map mechanical properties with a sub-cell resolution. Here, we demonstrate an inverted pulsed opto-acoustic microscope (iPOM) that operates in the 10 to 100 GHz range. These frequencies allow mapping quantitatively cell structures as thin as 10 nm and resolving the fibrillar details of cells. Using this non-invasive all-optical system, we produce high-resolution images based on mechanical properties as the contrast mechanisms, and we can observe the stiffness and adhesion of single migrating stem cells. The technique should allow transferring the diagnostic and imaging abilities of ultrasonic imaging to the single-cell scale, thus opening new avenues for cell biology and biomaterial sciences.

Scaled behavior of interface waves at an imperfect solid-solid interface

Laser ultrasonic techniques allow the remote analysis of adhesion mechanisms at imperfect interfaces up to GHz frequencies. However, the sensitivity of interface waves to the properties of the contact is not very well known. In the present work, the mechanical boundary conditions are described considering that the contacting solid half-spaces are connected by tangential and normal springs. Such a modeling implies a discontinuity of the displacement field across the interface. To identify the relative amplitudes of the different types of interface waves—skimming, leaky Rayleigh (LR) and Stoneley (St) waves—a semi-analytical time domain model describing the thermoelastic laser generation is derived. The results illustrate the influence of the boundary conditions on the attenuation of the LR wave and on the existence of the St wave. In addition, a single compact and elegant dispersion equation is presented to investigate the behaviour of the interface waves propagating along a generalized imperfect boundary....

Non-invasive optoacoustic probing of the density and stiffness of single biological cells

Recently, the coherent generation of GHz acoustic waves using ultrashort laser pulses has demonstrated the ability to probe the sound velocity in vegetal cells and in cell-mimicking soft micro-objects with micrometer resolution, opening tremendous potentialities for single-cell biology. However, manipulating biological media in physiological conditions is often a technical challenge when using a laser-based setup. In this article, we present a new opto-acoustic bio-transducer composed of a thin metal film sputtered on a transparent heat sink that allows reducing importantly the laser-induced cellular stresses, and offers a wide variety of optical configurations. In particular, by exploiting the acoustic reflection coefficient at the sample-transducer interface and the photoacoustic interaction inside the transparent sample, the density and compressibility of the sample can be probed simultaneously. Using an ad hoc signal analysis based on Hilbert and wavelet transforms, these quantities are measured accur...

Probing single-cell mechanics with picosecond ultrasonics

The mechanical properties of cells play a key role in several fundamental biological processes, such as migration, proliferation, differentiation and tissue morphogenesis. The complexity of the inner cell composition and the intricate meshwork formed by transmembrane cell-substrate interactions demands a non-invasive technique to probe cell mechanics and cell adhesion at a subcell scale. In this paper we review the use of laser-generated GHz acoustic waves--a technique called picosecond ultrasonics (PU)--to probe the mechanical properties of single cells. We first describe applications to vegetal cells and biomimetic systems. We show how these systems can be used as simple models to understand more complex animal cells. We then present an opto-acoustic bio-transducer designed for in vivo measurements in physiological conditions. We illustrate the use of this transducer through the simultaneous probing of the density and compressibility of Allium cepa cells. Finally, we demonstrate that this technique can quantify animal-cell adhesion on metallic surfaces by analyzing the acoustic pulses reflected off the cell-metal interface. This innovative approach allows investigating quantitatively cell mechanics without fluorescent labels or mechanical contact to the cell.

Vibrations of microspheres probed with ultrashort optical pulses

We use ultrashort optical pulses to excite and detect vibrations of single silica spheres with a diameter of 5 microm placed at the surface of an acoustically mismatched substrate. In addition to the photoelastic detection of picosecond longitudinal acoustic pulses propagating inside the bulk, we detect gigahertz acoustic resonances of the sphere through probe beam defocusing. The mode frequencies are in close accord with those calculated from the elastic vibrations of a free sphere. We also record a resonant enhancement in the amplitude of specific modes of two touching spheres.

Effect of laser pulse duration in picosecond ultrasonics

An optical grating has been introduced in a picosecond ultrasonics experiment, in order to vary continuously the duration of the laser beam pulse from 0.1to150ps. The evolution of the measured signal has been observed and analyzed through the comparison with a theoretical approach based on a two-temperature model. The latter allows matching the acoustic echoes together with the thermal background and the coincidence peak, for each pulse duration and at any time scale. The broadening of the acoustic echoes and the disappearing of its Brillouin component, along with the diminishing of the thermal coincidence peak, have been demonstrated when increasing the pulse duration. For a constant incident pulse energy, the efficiency of acoustic generation is optimum for the shortest pulses. Nevertheless, for longer pulses designed to obtain thermal conditions below the ablation threshold, acoustic generation could be enhanced.

Relaxation dynamics in single polymer microcapsules probed with laser-generated GHz acoustic waves

Using an ultrafast optical technique we probe coherent phonon propagation inside a single microcapsule composed of a nanometric polymer shell made of poly(lactide-co-glycolide) (PLGA) encapsulating a liquid perfluorooctyl bromide (PFOB) core. Longitudinal storage and loss moduli are measured simultaneously in the transparent shell and core at frequencies ∼18 and ∼4 GHz, respectively, using time-resolved Brillouin spectroscopy. A time-frequency analysis allows determination of the thicknesses of several capsule shells ranging from 620 down to 80 nm. Comparison with lower frequency data shows a weak power-law frequency dependence of phonon attenuation in the PLGA shell, the signature of thermally activated processes in glasses.

Optical tracking of picosecond coherent phonon pulse focusing inside a sub-micron object

By means of an ultrafast optical technique, we track focused gigahertz coherent phonon pulses in objects down to sub-micron in size. Infrared light pulses illuminating the surface of a single metal-coated silica fibre generate longitudinal-phonon wave packets. Reflection of visible probe light pulses from the fibre surface allows the vibrational modes of the fibre to be detected, and Brillouin optical scattering of partially transmitted light pulses allows the acoustic wavefronts inside the transparent fibre to be continuously monitored. We thereby probe acoustic focusing in the time domain resulting from generation at the curved fibre surface. An analytical model, supported by three-dimensional simulations, suggests that we have followed the focusing of the acoustic beam down to a ~150-nm diameter waist inside the fibre. This work significantly narrows the lateral resolution for focusing of picosecond acoustic pulses, normally limited by the diffraction limit of focused optical pulses to ~1 μm, and thereby opens up a new range of possibilities including nanoscale acoustic microscopy and nanoscale computed tomography.