Large colloidal probes for atomic force microscopy: Fabrication and calibration issues

Matteo Chighizola, Luca Puricelli, Ludovic Bellon, Alessandro Podestà, Journal of Molecular Recognition  e2849 (2020)

doi: 10.1002/jmr.2879

Atomic force microscopy (AFM) is a powerful tool to investigate interaction forces at the micro and nanoscale. Cantilever stiffness, dimensions and geometry of the tip can be chosen according to the requirements of the specific application, in terms of spatial resolution and force sensitivity. Colloidal probes (CPs), obtained by attaching a spherical particle to a tipless (TL) cantilever, offer several advantages for accurate force measurements: tunable and well-characterisable radius; higher averaging capa- bilities (at the expense of spatial resolution) and sensitivity to weak interactions; a well-defined interaction geometry (sphere on flat), which allows accurate and reliable data fitting by means of analytical models. The dynamics of standard AFM probes has been widely investigated, and protocols have been developed for the calibration of the cantilever spring constant. Nevertheless, the dynamics of CPs, and in particular of large CPs, with radius well above 10 μm and mass comparable, or larger, than the cantilever mass, is at present still poorly characterized. Here we describe the fabrica- tion and calibration of (large) CPs. We describe and discuss the peculiar dynamical behaviour of CPs, and present an alternative protocol for the accurate calibration of the spring constant.

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Calibrated force measurement in Atomic Force Microscopy using the Transient Fluctuation Theorem

Samuel Albert, Aubin Archambault, Artyom Petrosyan, Caroline Crauste-Thibierge, Ludovic Bellon, Sergio Ciliberto, EPL 131, 10008 (2020)

doi: 10.1209/0295-5075/131/10008

The Transient Fluctuation Theorem is used to calibrate an Atomic Force Microscope by measuring the fluctuations of the work performed by a time dependent force applied between a colloïdal probe and the surface. From this measure one can easily extract the value of the interaction force and the relevant parameters of the cantilever. The results of this analysis are compared with those obtained by standard calibration methods. 

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Extended equipartition in a mechanical system subject to a heat flow: the case of localised dissipation

Alex Fontana, Richard Pedurand, Ludovic Bellon, J. Stat. Mech. 073206 (2020)

doi:10.1088/1742-5468/ab97b1

Statistical physics in equilibrium grants us one of its most powerful tools: the equipartition principle. It states that the degrees of freedom of a mechanical system act as a thermometer: temperature is equal to the mean variance of their oscillations divided by their stiffness. However, when a non-equilibrium state is considered, this principle is no longer valid. In our experiment, we study the fluctuations of a micro-cantilever subject to a strong heat flow, which creates a highly non-uniform local temperature. We measure independently the temperature profile of the object and the temperature yielded from the mechanical thermometers, thus testing the validity of the equipartition principle out of equilibrium. We demonstrate how the fluctuations of the most energetic degrees of freedom are equivalent to the temperature at the base of the cantilever, even when the average temperature is several hundreds of degrees higher. We then present a model based on the localised mechanical dissipation in the system to account for our results, which correspond to mechanical losses localised at the clamping position.

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Extended Nyquist formula for a resistance subject to a heat flow

Benjamin Monnet, Sergio Ciliberto and Ludovic Bellon, J. Stat. Mech. 104011 (2019)

doi:10.1088/1742-5468/ab363c

The Nyquist formula quantifies the thermal noise driven fluctuations of voltage across a resistance in equilibrium. We deal here with the case of a resistance driven out of equilibrium by putting it in contact with two thermostats at different temperatures. We reach a non-equilibrium steady state where a heat flux is flowing through the resistance. Our measurements demonstrate anyway that a simple extension of the Nyquist formula to the non uniform temperature field describes with an excellent precision the thermal noise. For a metallic ohmic material, the fluctuations are actually equivalent to those of a resistance in equilibrium with a single thermostat at the mean temperature between the hot and cold sources. 

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Detecting protein folding by thermal fluctuations of microcantilevers

Romina Muñoz, Felipe Aguilar-Sandoval, Ludovic Bellon, and Francisco Melo, PLoS ONE 12, e0189979 (2017)

doi: 10.1371/journal.pone.0189979

The accurate characterization of proteins in both their native and denatured states is essential to effectively understand protein function, folding and stability. As a proof of concept, a micro rheological method is applied, based on the characterization of thermal fluctuations of a micro cantilever immersed in a bovine serum albumin solution, to assess changes in the viscosity associated with modifications in the protein’s structure under the denaturant effect of urea. Through modeling the power spectrum density of the cantilever’s fluctuations over a broad frequency band, it is possible to implement a fitting procedure to accurately determine the viscosity of the fluid, even at low volumes. Increases in viscosity during the denaturant process are identified using the assumption that the protein is a hard sphere, with a hydrodynamic radius that increases during unfolding. This is modeled accordingly through the Einstein-Batchelor formula. The Einstein-Batchelor formula estimates are verified through dynamic light scattering, which measures the hydrodynamic radius of proteins. Thus, this methodology is proven to be suitable for the study of protein folding in samples of small size at vanishing shear stresses.

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Low thermal fluctuations in a system heated out of equilibrium

Mickael Geitner, Felipe Aguilar Sandoval, Eric Bertin and Ludovic Bellon, Phys. Rev. E 95, 032138

doi: 10.1103/PhysRevE.95.032138

We study the mechanical fluctuations of a micrometer sized silicon cantilever subjected to a strong heat flow, thus having a highly non-uniform local temperature. In this non-equilibrium steady state, we show that fluctuations are equivalent to the thermal noise of a cantilever at equilibrium around room temperature, while its mean local temperature is several hundred of degrees higher. Changing the mechanical dissipation by adding a coating to the cantilever, we recover the expected rise of fluctuations with the mean temperature. Our work demonstrates that inhomogeneous dissipation mechanisms can decouple the amplitude of thermal fluctuations from the average temperature. This property could be useful to understand out-of-equilibrium fluctuating systems, or to engineer low noise instruments.

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Dynamic stiffness of the contact between a carbon nanotube and a flat substrate in a peeling geometry

Tianjun Li,  Lorène Champougny and Ludovic Bellon,  Journal of Applied Physics 121, 094305 (2017)

doi: 10.1063/1.4977867

We study the physics of adhesion and the contact mechanics at the nanoscale with a peeling experiment of a carbon nanotube on a flat substrate. Using an interferometric atomic force microscope and an extended force modulation protocol, we investigate the frequency response of the stiffness of the nano-contact from DC to 20 kHz. We show that this dynamic stiffness is only weakly frequency dependent, increasing by a factor 2 when the frequency grows by 3 orders of magnitude. Such behavior may be the signature of amorphous relaxations during the mechanical solicitation at the nano-scale.

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«Noiseless» thermal noise measurement of atomic force microscopy cantilevers

Basile Pottier and Ludovic Bellon, Appl. Phys. Lett. 110, 094105 (2017)

doi: 10.1063/1.4977790

When measuring quadratic values representative of random fluctuations, such as the thermal noise of Atomic Force Microscopy (AFM) cantilevers, the background measurement noise cannot be averaged to zero. We present a signal processing method that allows to get rid of this limitation using the ubiquitous optical beam deflection sensor of standard AFMs. We demonstrate a two orders of magnitude enhancement of the signal to noise ratio in our experiment, allowing the calibration of stiff cantilevers or easy identification of higher order modes from thermal noise measurements.

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High Resolution Viscosity Measurement by Thermal Noise Detection

Felipe Aguilar Sandoval, Manuel Sepúlveda, Ludovic Bellon, and Francisco Melo, Sensors 2015, 27905-27916 (2015)

doi: 10.3390/s151127905

An interferometric method is implemented in order to accurately assess the thermal fluctuations of a micro-cantilever sensor in liquid environments. The power spectrum density (PSD) of thermal fluctuations together with Sader’s model of the cantilever allow for the indirect measurement of the liquid viscosity with good accuracy. The good quality of the deflection signal and the characteristic low noise of the instrument allow for the detection and corrections of drawbacks due to both the cantilever shape irregularities and the uncertainties on the position of the laser spot at the fluctuating end of the cantilever. Variation of viscosity below 0.03 mPa·s was detected with the alternative to achieve measurements with a volume as low as 50 μL.

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