Jérôme Polesel-Maris

FluidFM: Combining Atomic Force Microscopy and Nanofluidics in a Universal Liquid Delivery System for Single Cell Applications and Beyond

We describe the fluidFM, an atomic force microscope (AFM) based on hollow cantilevers for local liquid dispensing and stimulation of single living cells under physiological conditions. A nanofluidic channel in the cantilever allows soluble molecules to be dispensed through a submicrometer aperture in the AFM tip. The sensitive AFM force feedback allows controlled approach of the tip to a sample for extremely local modification of surfaces in liquid environments. It also allows reliable discrimination between gentle contact with a cell membrane or its perforation. Using these two procedures, dyes have been introduced into individual living cells and even selected subcellular structures of these cells. The universality and versatility of the fluidFM will stimulate original experiments at the submicrometer scale not only in biology but also in physics, chemistry, and material science.

Rapid and Serial Quantification of Adhesion Forces of Yeast and Mammalian Cells

Cell adhesion to surfaces represents the basis for niche colonization and survival. Here we establish serial quantification of adhesion forces of different cell types using a single probe. The pace of single-cell force-spectroscopy was accelerated to up to 200 yeast and 20 mammalian cells per probe when replacing the conventional cell trapping cantilever chemistry of atomic force microscopy by underpressure immobilization with fluidic force microscopy (FluidFM). In consequence, statistically relevant data could be recorded in a rapid manner, the spectrum of examinable cells was enlarged, and the cell physiology preserved until approached for force spectroscopy. Adhesion forces of Candida albicans increased from below 4 up to 16 nN at 37°C on hydrophobic surfaces, whereas a Δhgc1-mutant showed forces consistently below 4 nN. Monitoring adhesion of mammalian cells revealed mean adhesion forces of 600 nN of HeLa cells on fibronectin and were one order of magnitude higher than those observed for HEK cells.

Parallel AFM imaging and force spectroscopy using two‐dimensional probe arrays for applications in cell biology

Atomic force microscopy (AFM) investigations of living cells provide new information in both biology and medicine. However, slow cell dynamics and the need for statistically significant sample sizes mean that data collection can be an extremely lengthy process. We address this problem by parallelizing AFM experiments using a two‐dimensional cantilever array, instead of a single cantilever. We have developed an instrument able to operate a two‐dimensional cantilever array, to perform topographical and mechanical investigations in both air and liquid. Deflection readout for all cantilevers of the probe array is performed in parallel and online by interferometry. Probe arrays were microfabricated in silicon nitride. Proof‐of‐concept has been demonstrated by analyzing the topography of hard surfaces and fixed cells in parallel, and by performing parallel force spectroscopy on living cells. These results open new research opportunities in cell biology by measuring the adhesion and elastic properties of a large number of cells. Both properties are essential parameters for research in metastatic cancer development. Copyright

A tuning fork based wide range mechanical characterization tool with nanorobotic manipulators inside a scanning electron microscope

This study proposes a tuning fork probe based nanomanipulation robotic system for mechanical characterization of ultraflexible nanostructures under scanning electron microscope. The force gradient is measured via the frequency modulation of a quartz tuning fork and two nanomanipulators are used for manipulation of the nanostructures. Two techniques are proposed for attaching the nanostructure to the tip of the tuning fork probe. The first technique involves gluing the nanostructure for full range characterization whereas the second technique uses van der Waals and electrostatic forces in order to avoid destroying the nanostructure. Helical nanobelts (HNB) are proposed for the demonstration of the setup. The nonlinear stiffness behavior of HNBs during their full range tensile studies is clearly revealed for the first time. Using the first technique, this was between 0.009 N/m for rest position and 0.297 N/m before breaking of the HNB with a resolution of 0.0031 N/m. For the second experiment, this was between 0.014 N/m for rest position and 0.378 N/m before detaching of the HNB with a resolution of 0.0006 N/m. This shows the wide range sensing of the system for potential applications in mechanical property characterization of ultraflexible nanostructures.

A virtual dynamic atomic force microscope for image calculations

Calculations of frequency modulation-atomic force microscopy (FM-AFM) images are presented. A virtual FM-AFM, which realistically simulates the experiment by including the control system of the microscope, is implemented in order to go beyond the usual static approximation. It is shown that the results obtained within the static approach can be recovered in the limit of small scanning speed, while images at realistic scanning speed are distorted. The influence of the experimental noise on the images is investigated, allowing us to evaluate the sensitivity of the instrument.

Piezoresistive cantilever array for life sciences applications

Atomic Force Microscopy (AFM) techniques are used with one- or two-dimensional arrays of piezoresistive probes for parallel imaging. We present a newly designed AFM platform to drive these passivated piezoresistive cantilever arrays in air and liquid environments. Large area imaging in liquid as well as qualitative and quantitative analysis of biological cells are demonstrated by the means of piezoresistive cantilever for the first time to our knowledge. Noise limitations in topography and force resolutions of these piezolevers are quantified.

Nanostenciling for fabrication and interconnection of nanopatterns and microelectrodes

Stencil lithography is used for patterning and connecting nanostructures with metallic microelectrodes in ultrahigh vacuum. Microelectrodes are fabricated by static stencil deposition through a thin silicon nitride membrane. Arbitrary nanoscale patterns are then deposited at a predefined position relative to the microelectrodes, using as a movable stencil mask an atomic force microscopy (AFM) cantilever in which apertures have been drilled by focused ion beam. Large scale AFM imaging, combined with the use of a high precision positioning table, allows inspecting the microelectrodes and positioning the nanoscale pattern with accuracy better than 100nm.

Combined dynamic scanning tunneling microscopy and frequency modulation atomic force microscopy investigations on polythiophene chains on graphite with a tuning fork sensor

Polythiophene molecules adsorbed on a highly oriented pyrolytic graphite surface were studied by combined dynamic scanning tunneling microscopy (STM) and frequency modulation atomic force microscopy (FM-AFM) with a quartz tuning fork sensor operating in Qplus mode and equipped with a Pt/Ir tip. Upon completing a careful sub-angstrom oscillation amplitude calibration of the probe, experiments were conducted in an ultra high vacuum at room temperature. By selecting the tip/surface distance regulation parameter, one can select the type of simultaneous information obtained in an area. For distance regulation based on the mean tunneling current, dynamic STM images together with maps of tip/surface force gradient were obtained. FM-AFM images with maps of the tunneling current were also acquired when the distance regulation was based on the frequency shift. Comparison between these images reveals interesting features. For example the tip which operates in STM mode with ultra low current (<10 pA) generates differ...

Experimental three-dimensional description of the liquid hexadecane/graphite interface.

By using an atomic force microscope based on a quartz tuning fork sensor, a 3-dimensional description of the interface between liquid hexadecane and a highly oriented pyrolytic graphite surface can be achieved at room temperature. The C16H34 monolayer in contact with the substrate surface exhibits a lamellar structure whereas no observation at the liquid/graphite interface by scanning tunnelling microscopy was reported for this alkane. The second layer shows very weak corrugations corresponding to lamella boundaries. Force/distance curves show at least four oscillations separated by 0.4 nm except for the first period with a 0.38 nm distance that corresponds to the layer closer the substrate. Such a description agrees well with molecular dynamics results obtained on alkane/solid interfaces.

Is atomic-scale dissipation in NC-AFM real? Investigation using virtual atomic force microscopy

Using a virtual dynamic atomic force microscope, that explicitly simulates the operation of a non-contact AFM experiment, we have performed calculations to investigate the formation of atomic-scale contrast in dissipation images. A non-conservative tip?surface interaction was implemented using the theory of dynamical response in scanning probe microscopy with energies and barriers derived from realistic atomistic modelling. It is shown how contrast in the damping signal is due to the hysteresis in the tip?surface force and not an artefact of the finite response of the complicated instrumentation. Topography and dissipation images of the CaO(001) surface are produced which show atomic-scale contrast in the dissipation with a corrugation of approximately 0.1?eV, which is typical of that observed in images of similar binary ionic surfaces. The effect of the fast-direction scanning speed on the image formation is also investigated and discussed.