Nanoscale bioelectrical characterization

Gabriel Gomila Lluch | Group Leader
Ricardo Hidalgo Gonzalez | Postdoctoral Researcher
Lázaro René Izquierdo Fábregas | Postdoctoral Researcher
Adrica Kyndiah | Postdoctoral Researcher
Martí Checa Nualart | PhD Student
Martina Di Muzio | PhD Student
Helena Lozano Caballero | PhD Student
Rubén Millán Solsona | Laboratory Technician


The main goal of the Nanoscale Bioelectrical Characterization group is to develop new experimental setups based on atomic force microscopy and theoretical frameworks enabling the access to the electrical properties of biological systems at the nanoscale (including biomembranes, single viruses, single bacteria cells and eukaryotic cells).

Our main objective is to contribute to develop new label-free biological nanoscale characterization methods and new electronic biosensors.

Above right: top row: Topographic images of a single bacterial endospore measured by Atomic Force Microscopy at different environmental relative humidity conditions. Bottom Row: Dielectric images of the same bacterial endospore measured by Electrostatic Force Microscopy under the same relative humidity conditions. While the topography of the bacterial endospore remains almost unaltered when modifying the environmental relative humidity, the dielectric response changes dramatically. From these type of measurements, the internal hydration properties of a single endospore can be obtained.

During 2016 we have determined, for the first time, the electromagnetic properties of single bacteria cells in the high frequency range (> GHz) with the use of the Scanning Microwave Microscope and of specific 3D numerical simulation models. We showed that with this approach one can detect the presence of small-scale nanostructures inside microorganisms, providing endless applications in the label-free imaging of single bacterial cells at high spatial resolution.

On the other side, we have probed the internal hydration properties of single bacterial endospores by means of Electrostatic Force Microscopy. Endospores are recognized as the hardiest form of life on Earth, and one of the reasons for this, is that they handle changes in environmental relative humidity in a very smart way.

Left: Atomic Force Microscopy topographic image of embryonic mouse cortical neurons fixed with paraformaldehyde and imaged in air in intermittent contact mode (sample provided by the group of Prof. J. A. del Rio).

We have shown in a label-free way that the endospores are able to preserve their core, where DNA is located, under low hydration conditions, what is key to understand the endospore’s extraordinary survival abilities. On the methodological aspects, we have continued our efforts towards providing a simple interpretation to Electrostatic Force Microscopy and Scanning Microwave Microscopy images of highly non-planar samples, such as single bacterial or eukaryotic cells, for which we have developed a method to remove topographic cross-talk effects from the images.

Finally, we have also optimized sample preparation and imaging methods to image and study electrically excitable cells with the Atomic Force Microscope, such as neurons.

Right: Electrical potential distribution corresponding to the electric interaction between a voltage biased sharp conducting tip of radius 250 nm and a single bacterial cell. The bacterial cell is represented as a 3D ellipsoid structure with uniform electric polarization. From the calculated electric potential distribution the tip-bacteria capacitance can be calculated and compared to experimental measurements obtained with the Scanning Microwave Microscope, in order to determine the electric permittivity of a single bacteria cell at GHz frequencies.


Training the next generation of advanced microscopy experts

The Nanoscale Biolectrical Characterization group has been awarded EU funding to coordinate a project that aims to train a new generation of researchers in the science and technology of Scanning Probe Microscopes.

Marie Skłodowska-Curie Early Stage Researcher (PhD student) on Nanoscale Tomography based on Electrostatic Force Microscopy
Application Deadline: 28/02/2017

The Nanoscale Biolectrical Characterization group is looking for a Early Stage Researcher (PhD student) to develop his/her PhD thesis project on the development of a novel Nano-tomographic technique based on electrostatic force microscopy.

Marie Skłodowska-Curie Early Stage Researcher (PhD student) on Nanoscale Composition Mapping with Electrostatic Force Microscopy
Application Deadline: 28/02/2017

The Nanoscale Biolectrical Characterization group is looking for a Early Stage Researcher (PhD student) to develop his/her PhD thesis project on the label-free mapping of biological membranes’ composition with nanoscale spatial resolution.

Using EFM to probe the secrets of bacterial endospore survival strategies

An IBEC group has demonstrated, for the first time, that the hydration properties of a single bacterial endospore in varying environmental relative humidity can be determined with high accuracy and reproducibility, and in a non-destructive way, shedding new light on endospore survival strategies.

Microwave electromagnetic properties of single bacterial cells measured for the first time

Researchers at IBEC and their collaborators from the Johannes Kepler University of Linz, The University of Manchester and the company Keysight Technologies have now achieved an elusive goal: to measure the electromagnetic properties of biological materials at the level of a single bacterial cell and at very high frequencies (gigahertz).

Another big step towards understanding the electric properties of the cell

Having measured the electric polarizability of DNA – a fundamental property that directly influences its biological functions – for the first time ever last year, IBEC´s Nanoscale Bioelectrical Characterization group has made a further breakthrough in the understanding of the dielectric properties of cell constituents by measuring the electric polarizability of the main components of the cell membrane – namely lipids, sterols and proteins – with a spatial resolution down to 50nm.

ICREA Academia Award for IBEC group leader

Gabriel Gomila, IBEC group leader and Associate Professor at the UB, has received an ICREA Academia Prize 2014 for excellence in research and capacity for leadership.

IBEC internal collaboration succeeds in measuring bacterial cell response to electrical fields

Two groups working together at IBEC demonstrate the potential of electrical studies of single bacterial cells in a paper published in ACS Nano. Gabriel Gomila’s Nanoscale Bioelectrical Characterization group and that of Antonio Juárez, Microbial Biotechnology and Host-pathogen Interaction, combined their expertise on microscopic electrical measurements and bacteria respectively to come up with a way to study the response to external electrical fields of just a single bacterial cell.

Researchers measure a property of DNA for the first time

The electric polarizability of DNA is a fundamental property that directly influences its biological functions. Despite the importance of this property, however, its measurement has remained elusive so far. In a study published in PNAS today, researchers at Barcelona’s Institute for Bioengineering of Catalonia (IBEC) led by Laura Fumagalli, senior researcher at IBEC and lecturer at the University of Barcelona, and their collaborators at the Institute for Research in Biomedicine (IRB) and at Barcelona Supercomputing Center (BSC), and at Centro Nacional de Biotecnologia (CNB-CSIC) and IMDEA Nanociencia in Madrid, describe how they have found a way to directly measure DNA electric polarizability – represented by its dielectric constant, which indicates how a material reacts to an applied electric field – for the first time ever.

IBEC research on the cover of Nanotechnology

The latest article published by IBEC’s Nanoscale bioelectrical characterization group has made the cover of the journal Nanotechnology.

“Nanomicrowave” cooking up something new

A new European Marie Curie Initial Training Network involving IBEC’s Nanoscale Bioelectrical Characterization group will attempt to bring research into microwaves – which are extensively used in a host of applications such as telecommunications, microwave ovens and radar – to a whole new level.

‘Fingerprinting’ nanoscale objects and viruses

Scientists at IBEC in Barcelona have found a way of effectively identifying nanoscale objects and viruses that could offer a breakthrough for biomedical diagnostics, environmental protection and nano-electronics.

Microscope development project for sub-surface imaging at the nanoscale

IBEC’s Nanoscale Bioelectrical Characterization group, headed by Gabriel Gomila, is a partner in a new EU-funded collaborative project set to develop a new tool for non-destructive 3D nanoscale structural characterization, the Volumetric Scanning Microwave Microscope (VSMM).

New technique to study the electrical activity of the cellular membrane

Gabriel Gomila and Laura Fumagalli, from the Nanoscale bioelectrical characterization line at IBEC, are two of the authors of the study.


EU-funded projects
Scanning probe microscopies for nanoscale fast, tomographic and composition imaging (SPM2.0) (2017-2020) Marie Curie Skłodowska European Training Network (MSCA-ITN-ETN) Gabriel Gomila (Project Coordinator)
BIOWIRESENSE Plataforma universal para la detección de biomarcadores basada en nanocables bacterianos conductores (2017-2019) MINECO, Explora Ciencia Gabriel Gomila
National projects
NANOELECTOMOGRAPHY Electrical nanotomography based on scanning probe microscopy for nanomaterials and biological samples (2014-2016) MINECO (TEC2013-48344-C2-1-P) Gabriel Gomila
NANOELECTROPHYS Scanning Electric Force Microscope for Electrophysological Recordings at the Nanoscale
MINECO (TEC2016-79156-P) Gabriel Gomila
ICREA Academia Award (2015-2019) Catalan Institution for Research and Advanced Studies (ICREA) / Generalitat de Catalunya Gabriel Gomila
Finished projects
NANOMICROWAVE Microwave Nanotechnology for Semiconductor and Life Sciences (2013-2016) MARIE CURIE – ITN Gabriel Gomila
V-SMMART Nano Volumetric Scanning Microwave Microscopy Analytical and Research Tool for Nanotechnology (2012-2016) NMP – SME Gabriel Gomila
AFM4NanoMed&Bio European network on applications of Atomic Force Microscopy to Nanomedicine and Life Sciences EU COST Action TD1002 Gabriel Gomila (Management Committee Substitute Member)


Biagi, Maria Chiara, Badino, Giorgio, Fabregas, Rene, Gramse, Georg, Fumagalli, Laura, Gomila, Gabriel, (2017). Direct mapping of the electric permittivity of heterogeneous non-planar thin films at gigahertz frequencies by scanning microwave microscopy Physical Chemistry Chemical Physics 19, (5), 3884-3893

We obtained maps of the electric permittivity at ~19 GHz frequencies on non-planar thin film heterogeneous samples by means of combined atomic force-scanning microwave microscopy (AFM-SMM). We show that the electric permittivity maps can be obtained directly from the capacitance images acquired in contact mode, after removing the topographic cross-talk effects. This result demonstrates the possibility to identify the electric permittivity of different materials in a thin film sample irrespectively of their thickness by just direct imaging and processing. We show, in addition, that quantitative maps of the electric permittivity can be obtained with no need of any theoretical calculation or complex quantification procedure when the electric permittivity of one of the materials is known. To achieve these results the use of contact mode imaging is a key factor. For non-contact imaging modes the effects of the local sample thickness and of the imaging distance makes the interpretation of the capacitance images in terms of the electric permittivity properties of the materials much more complex. Present results represent a substantial contribution to the field of nanoscale microwave dielectric characterization of thin film materials with important implications for the characterization of novel 3D electronic devices and 3D nanomaterials.

Van Der Hofstadt, Marc, Fabregas, Rene, Millan, Ruben, Juarez, Antonio, Fumagalli, Laura, Gomila, Gabriel, (2016). Internal hydration properties of single bacterial endospores probed by electrostatic force microscopy ACS Nano 10, (12), 11327–11336

We show that the internal hydration properties of single Bacillus cereus endospores in air under different relative humidity (RH) conditions can be determined through the measurement of its electric permittivity by means of quantitative electrostatic force microscopy (EFM). We show that an increase in the RH from 0% to 80% induces a large increase in the equivalent homogeneous relative electric permittivity of the bacterial endospores, from ~4 up to ~17, accompanied only by a small increase in the endospore height, of just a few nanometers. These results correlate the increase of the moisture content of the endospore with the corresponding increase of environmental RH. 3D finite element numerical calculations, which include the internal structure of the endospores, indicate that the moisture is mainly accumulated in the external layers of the endospore, hence preserving the core of the endospore at low hydration levels. This mechanism is different from what we observe for vegetative bacterial cells of the same species, in which the cell wall at high humid atmospheric conditions is not able to preserve the cytoplasmic region at low hydration levels. These results show the potential of quantitative EFM under environmental humidity control to study the hygroscopic properties of small scale biological (and non-biological) entities and to determine its internal hydration state. A better understanding of nano-hygroscopic properties can be of relevance in the study of essential biological processes and in the design of bio-nanotechnological applications.

Biagi, Maria Chiara, Fabregas, Rene, Gramse, Georg, Van Der Hofstadt, Marc, Juárez, Antonio, Kienberger, Ferry, Fumagalli, Laura, Gomila, Gabriel, (2016). Nanoscale electric permittivity of single bacterial cells at gigahertz frequencies by scanning microwave microscopy ACS Nano 10, (1), 280-288

We quantified the electric permittivity of single bacterial cells at microwave frequencies and nanoscale spatial resolution by means of near-field scanning microwave microscopy. To this end, calibrated complex admittance images have been obtained at ?19 GHz and analyzed with a methodology that removes the nonlocal topographic cross-talk contributions and thus provides quantifiable intrinsic dielectric images of the bacterial cells. Results for single Escherichia coli cells provide a relative electric permittivity of ?4 in dry conditions and ?20 in humid conditions, with no significant loss contributions. Present findings, together with the ability of microwaves to penetrate the cell membrane, open an important avenue in the microwave label-free imaging of single cells with nanoscale spatial resolution.

Van Der Hofstadt, M., Fabregas, R., Biagi, M.C., Fumagalli, L., Gomila, G., (2016). Nanoscale dielectric microscopy of non-planar samples by lift-mode electrostatic force microscopy Nanotechnology 27, (40), 405706

Lift-mode electrostatic force microscopy (EFM) is one of the most convenient imaging modes to study the local dielectric properties of non-planar samples. Here we present the quantitative analysis of this imaging mode. We introduce a method to quantify and subtract the topographic crosstalk from the lift-mode EFM images, and a 3D numerical approach that allows for extracting the local dielectric constant with nanoscale spatial resolution free from topographic artifacts. We demonstrate this procedure by measuring the dielectric properties of micropatterned SiO 2 pillars and of single bacteria cells, thus illustrating the wide applicability of our approach from materials science to biology.

Dols-Perez, Aurora, Gramse, Georg, Calo, Annalisa, Gomila, Gabriel, Fumagalli, Laura, (2015). Nanoscale electric polarizability of ultrathin biolayers on insulator substrates by electrostatic force microscopy Nanoscale 7, 18327-18336

We measured and quantified the local electric polarization properties of ultrathin (~ 5 nm) biolayers on mm-thick mica substrates. We achieved it by scanning a sharp conductive tip (< 10 nm radius) of an electrostatic force microscope over the biolayers and quantifying sub-picoNewton electric polarization forces with a sharp-tip model implemented using finite-element numerical calculations. We obtained relative dielectric constants ?r = 3.3, 2.4 and 1.9 for bacteriorhodopsin, dioleoylphosphatidylcholine (DOPC) and cholesterol layers, chosen as representative of the main cell membrane components, with an error below 10% and a spatial resolution down to ~ 50 nm. The ability of using insulating substrates common in biophysics research, such as mica or glass, instead of metallic substrates, offers both a general platform to determine the dielectric properties of biolayers and a wider compatibility with other characterization techniques, such as optical microscopy. This opens up new possibilities for biolayer research at the nanoscale, including nanoscale label-free composition mapping.

Van Der Hofstadt, M., Hüttener, M., Juárez, A., Gomila, G., (2015). Nanoscale imaging of the growth and division of bacterial cells on planar substrates with the atomic force microscope Ultramicroscopy 154, 29-36

Abstract With the use of the atomic force microscope (AFM), the Nanomicrobiology field has advanced drastically. Due to the complexity of imaging living bacterial processes in their natural growing environments, improvements have come to a standstill. Here we show the in situ nanoscale imaging of the growth and division of single bacterial cells on planar substrates with the atomic force microscope. To achieve this, we minimized the lateral shear forces responsible for the detachment of weakly adsorbed bacteria on planar substrates with the use of the so called dynamic jumping mode with very soft cantilever probes. With this approach, gentle imaging conditions can be maintained for long periods of time, enabling the continuous imaging of the bacterial cell growth and division, even on planar substrates. Present results offer the possibility to observe living processes of untrapped bacteria weakly attached to planar substrates.

Keywords: Atomic Force Microscope (AFM), Living cell imaging, Bacteria division, Gelatine immobilization, Dynamic jumping mode

Botaya, Luis, Otero, Jorge, González, Laura, Coromina, Xavier, Gomila, Gabriel, Puig-Vidal, Manel, (2015). Quartz tuning fork-based conductive atomic force microscope with glue-free solid metallic tips Sensors and Actuators A: Physical 232, 259-266

Abstract Here, we devise a conductive Atomic Force Microscope (C-AFM) based on quartz tuning forks (QTFs) and metallic tips capable of simultaneously imaging the topography and conductance of a sample with nanoscale spatial resolution. The system is based on a header design which allows the metallic tip to be placed in tight and stable mechanical contact with the QTF without the need to use any glue. This allows electrical measurements to be taken with an electrically excited QTF with the two prongs free. The amplitude oscillation of the QTF is used to control the tip-sample distance and to acquire the topographic images. Meanwhile, the metallic tip is connected to a current–voltage amplifier circuit to measure the tip-sample field emission/tunneling current and to produce the conductive images. This method allows decoupled electrical measurement of the topography and electrical properties of the sample. The results we obtain from calibration samples demonstrate the feasibility of this measurement method and the adequacy of the performance of the system.

Keywords: AFM, Conductive AFM, Quartz tuning fork

Esteban-Ferrer, Daniel, Edwards, Martin Andrew, Fumagalli, Laura, Juarez, Antonio, Gomila, Gabriel, (2014). Electric polarization properties of single bacteria measured with electrostatic force microscopy ACS Nano 8, (10), 9843–9849

We quantified the electrical polarization properties of single bacterial cells using electrostatic force microscopy. We found that the effective dielectric constant, εr,eff , for the four bacterial types investigated (Salmonella Typhimurium, Escherchia coli, Lactobacilus sakei and Listeria innocua) is around 3-5 under dry air conditions. Under ambient humidity, it increases to εr,eff~6-7 for the Gram-negative bacterial types (S. Typhimurium and E. coli) and to εr,eff~15-20 for the Gram-positive ones (L. sakei and L. innocua). We show that the measured effective dielectric constants can be consistently interpreted in terms of the electric polarization properties of the biochemical components of the bacterial cell compartments and of their hydration state. These results demonstrate the potential of electrical studies of single bacterial cells. We quantified the electrical polarization properties of single bacterial cells using electrostatic force microscopy. We found that the effective dielectric constant, εr,eff , for the four bacterial types investigated (Salmonella Typhimurium, Escherchia coli, Lactobacilus sakei and Listeria innocua) is around 3-5 under dry air conditions. Under ambient humidity, it increases to εr,eff~6-7 for the Gram-negative bacterial types (S. Typhimurium and E. coli) and to εr,eff~15-20 for the Gram-positive ones (L. sakei and L. innocua). We show that the measured effective dielectric constants can be consistently interpreted in terms of the electric polarization properties of the biochemical components of the bacterial cell compartments and of their hydration state. These results demonstrate the potential of electrical studies of single bacterial cells.

Cuervo, A., Dans, P. D., Carrascosa, J. L., Orozco, M., Gomila, G., Fumagalli, L., (2014). Direct measurement of the dielectric polarization properties of DNA Proceedings of the National Academy of Sciences of the United States of America 111, (35), E3624-E3630

The electric polarizability of DNA, represented by the dielectric constant, is a key intrinsic property that modulates DNA interaction with effector proteins. Surprisingly, it has so far remained unknown owing to the lack of experimental tools able to access it. Here, we experimentally resolved it by detecting the ultraweak polarization forces of DNA inside single T7 bacteriophages particles using electrostatic force microscopy. In contrast to the common assumption of low-polarizable behavior like proteins (εr ∼ 2-4), we found that the DNA dielectric constant is ∼8, considerably higher than the value of ∼3 found for capsid proteins. State-of-the-art molecular dynamic simulations confirm the experimental findings, which result in sensibly decreased DNA interaction free energy than normally predicted by Poisson-Boltzmann methods. Our findings reveal a property at the basis of DNA structure and functions that is needed for realistic theoretical descriptions, and illustrate the synergetic power of scanning probe microscopy and theoretical computation techniques.

Keywords: Atomic force microscopy, Atomistic simulations, DNA packaging, DNA-ligand binding, Poisson-Boltzmann equation, capsid protein, DNA, double stranded DNA, amino acid composition, article, atomic force microscopy, bacteriophage, bacteriophage T7, dielectric constant, dipole, DNA binding, DNA packaging, DNA structure, electron microscopy, ligand binding, nonhuman, polarization, priority journal, protein analysis, protein DNA interaction, scanning probe microscopy, static electricity, virion, virus capsid, virus particle, atomic force microscopy, atomistic simulations, DNA packaging, DNA-ligand binding, Poisson-Boltzmann equation, Bacteriophage T7, Capsid, Cations, Dielectric Spectroscopy, DNA, DNA, Viral, DNA-Binding Proteins, Electrochemical Techniques, Ligands, Microscopy, Atomic Force, Models, Chemical, Nuclear Proteins

Caló, A., Reguera, D., Oncins, G., Persuy, M. A., Sanz, G., Lobasso, S., Corcelli, A., Pajot-Augy, E., Gomila, G., (2014). Force measurements on natural membrane nanovesicles reveal a composition-independent, high Young's modulus Nanoscale 6, (4), 2275-2285

Mechanical properties of nano-sized vesicles made up of natural membranes are crucial to the development of stable, biocompatible nanocontainers with enhanced functional, recognition and sensing capabilities. Here we measure and compare the mechanical properties of plasma and inner membrane nanovesicles ∼80 nm in diameter obtained from disrupted yeast Saccharomyces cerevisiae cells. We provide evidence of a highly deformable behaviour for these vesicles, able to support repeated wall-to-wall compressions without irreversible deformations, accompanied by a noticeably high Young's modulus (∼300 MPa) compared to that obtained for reconstituted artificial liposomes of similar size and approaching that of some virus particles. Surprisingly enough, the results are approximately similar for plasma and inner membrane nanovesicles, in spite of their different lipid compositions, especially on what concerns the ergosterol content. These results point towards an important structural role of membrane proteins in the mechanical response of natural membrane vesicles and open the perspective to their potential use as robust nanocontainers for bioapplications.

Dols-Perez, A., Fumagalli, L., Gomila, G., (2014). Structural and nanomechanical effects of cholesterol in binary and ternary spin-coated single lipid bilayers in dry conditions Colloids and Surfaces B: Biointerfaces 116, 295-302

We investigate the effects of Cholesterol (Chol) in the structural and nanomechanical properties of binary and ternary spin-coated single lipid bilayers made of Dioleoylphosphatidylcholine (DOPC) and Sphingomyelin (SM) in dry conditions. We show that for the DOPC/Chol bilayers, Chol induces an initial increase of the bilayer thickness, followed by decrease for concentrations above 30% Chol. The mechanical properties, instead, appear practically insensitive to the Chol content. For the SM/Chol bilayers we have observed both the thinning of the bilayer and the decrease of the force necessary to break it for Chol content above 40. mol%. In both binary mixtures phase separation is not observed. For ternary single bilayers of DOPC/SM/Chol, Chol induces phase segregation and the formation of domains resembling lipid rafts. The domains show a thickness and mechanical response clearly distinct from the surrounding phase and dependent on the relative Chol content. Based on the results obtained for the binary mixtures, DOPC- and SM-enriched domains can be identified. We highlight that many of the effects of Chol reported here for the dry multicomponent single lipid bilayers resemble closely those observed in hydrated bilayers, thus offering an additional insight into their properties.

Keywords: AFM, Air-stable lipid layer, Force spectroscopy, Lipid raft, Spin-coating

Gramse, G., Kasper, M., Fumagalli, L., Gomila, G., Hinterdorfer, P., Kienberger, F., (2014). Calibrated complex impedance and permittivity measurements with scanning microwave microscopy Nanotechnology 25, (14), 145703 (8)

We present a procedure for calibrated complex impedance measurements and dielectric quantification with scanning microwave microscopy. The calibration procedure works in situ directly on the substrate with the specimen of interest and does not require any specific calibration sample. In the workflow tip-sample approach curves are used to extract calibrated complex impedance values and to convert measured S11 reflection signals into sample capacitance and resistance images. The dielectric constant of thin dielectric SiO2 films were determined from the capacitance images and approach curves using appropriate electrical tip-sample models and the εr value extracted at f = 19.81 GHz is in good agreement with the nominal value of εr ∼ 4. The capacitive and resistive material properties of a doped Si semiconductor sample were studied at different doping densities and tip-sample bias voltages. Following a simple serial model the capacitance-voltage spectroscopy curves are clearly related to the semiconductor depletion zone while the resistivity is rising with falling dopant density from 20 Ω to 20 kΩ. The proposed procedure of calibrated complex impedance measurements is simple and fast and the accuracy of the results is not affected by varying stray capacitances. It works for nanoscale samples on either fully dielectric or highly conductive substrates at frequencies between 1 and 20 GHz.

Keywords: Complex impedance, Dielectric constant, Nanotechnology: calibration, Resistivity, Scanning microwave microscopy

Gomila, G., Gramse, G., Fumagalli, L., (2014). Finite-size effects and analytical modeling of electrostatic force microscopy applied to dielectric films Nanotechnology 25, (25), 255702 (11)

A numerical analysis of the polarization force between a sharp conducting probe and a dielectric film of finite lateral dimensions on a metallic substrate is presented with the double objective of (i) determining the conditions under which the film can be approximated by a laterally infinite film and (ii) proposing an analytical model valid in this limit. We show that, for a given dielectric film, the critical diameter above which the film can be modeled as laterally infinite depends not only on the probe geometry, as expected, but mainly on the film thickness. In particular, for films with intermediate to large thicknesses (>100 nm), the critical diameter is nearly independent from the probe geometry and essentially depends on the film thickness and dielectric constant following a relatively simple phenomenological expression. For films that can be considered as laterally infinite, we propose a generalized analytical model valid in the thin-ultrathin limit (<20-50 nm) that reproduces the numerical calculations and the experimental data. Present results provide a general framework under which accurate quantification of electrostatic force microscopy measurements on dielectric films on metallic substrates can be achieved.

Keywords: Dielectric constant, Dielectric films, Electrostatic force microscopy, Quantification, Analytical models, Electric force microscopy, Electrostatic force, Film thickness, Permittivity, Probes, Substrates, Ultrathin films, Accurate quantifications, Electrostatic force microscopy, Finite size effect, Lateral dimension, Metallic substrate, Numerical calculation, Polarization forces, Quantification, Dielectric films

Fumagalli, L., Edwards, Martin Andrew, Gomila, G., (2014). Quantitative electrostatic force microscopy with sharp silicon tips Nanotechnology 25, (49), 495701 (9)

Electrostatic force microscopy (EFM) probes are typically coated in either metal (radius ~ 30 nm) or highly-doped diamond (radius ~ 100 nm). Highly-doped silicon probes, which offer a sharpened and stable tip apex (radius ~ 1–10 nm) and are usually used only in standard atomic force microscopy, have been recently shown to allow enhanced lateral resolution in quantitative EFM and its application for dielectric constant measurement. Here we present the theoretical modelling required to quantitatively interpret the electrostatic force between these sharpened tips and samples. In contrast to a sphere-capped cone geometry used to describe metal/diamond-coated tips, modelling a sharpened silicon tip requires a geometry comprised of a cone with two different angles. Theoretical results are supported by experimental measurements of metallic substrates and ~10 nm radius dielectric nanoparticles. This work is equally applicable to EFM and other electrical scanned probe techniques, where it allows quantifying electrical properties of nanomaterials and 3D nano-objects with higher resolution.

Keywords: AFM, Dielectric constant, EFM, Dielectrics, Nanoparticles, Sharp tips

Castillo-Fernandez, O., Rodriguez-Trujillo, R., Gomila, G., Samitier, J., (2014). High-speed counting and sizing of cells in an impedance flow microcytometer with compact electronic instrumentation Microfluidics and Nanofluidics 16, (1-2), 91-99

Here we describe a high-throughput impedance flow cytometer on a chip. This device was built using compact and inexpensive electronic instrumentation. The system was used to count and size a mixed cell sample containing red blood cells and white blood cells. It demonstrated a counting capacity of up to ~500 counts/s and was validated through a synchronised high-speed optical detection system. In addition, the device showed excellent discrimination performance under high-throughput conditions.

Keywords: Electronics, Impedance, Microcytometry, Microfluidics, Red blood cells (RBCs), White blood cells (WBCs)

Birhane, Y., Otero, J., Pérez-Murano, F., Fumagalli, L., Gomila, G., Bausells, J., (2014). Batch fabrication of insulated conductive scanning probe microscopy probes with reduced capacitive coupling Microelectronic Engineering 119, 44-47

We report a novel fabrication process for the batch fabrication of insulated conductive scanning probe microscopy (SPM) probes for electrical and topographic characterization of soft samples in liquid media at the nanoscale. The whole SPM probe structure is insulated with a dielectric material except at the very tip end and at the contact pad area to minimize the leakage current in liquid. Additionally, the geometry of the conducting layer in the probe cantilever and substrate is engineered to reduce the parasitic capacitance coupling with the sample. The electrical characterization of the probes has shown that parasitic capacitances are significantly reduced as compared to fully metallized cantilevers.

Keywords: Conductive scanning probe microscopy (C-SPM), EFM, SECM, SECM-AFM, SIM

Caballero, D., Fumagalli, L., Teixidor, F., Samitier, J., Errachid, A., (2013). Directing polypyrrole growth by chemical micropatterns: A study of high-throughput well-ordered arrays of conductive 3D microrings Sensors and Actuators B: Chemical 177, 1003-1009

An array of well-ordered conducting polypyrrole microrings doped with cobaltabisdicarbollide [Co(C2B9H11) 2]- anions was fabricated by means of electropolymerization and submerged micro-contact printing techniques. The different conductive properties of the micropatterned thiols acted as a template for directing the electrochemical 3D growth of the microstructures over large areas. X-ray photoelectron spectroscopy characterization confirmed the presence of this unusual doping anion within the polymer. Its intrinsic properties together with hydrophobic interactions with the thiols guided the formation of the ring structures. A topographic study by atomic force microscopy gave insights into the PPy/[Co(C2B9H11) 2]- growing mechanism which is in agreement with the theoretical model of metal growth. Finally, the conductive properties of the microstructures were addressed by conductive-atomic force microscopy, showing a highly conductive behaviour. This methodology using cobaltabisdicarbollide as dopant anion could have important applications in organic microelectronics for the development of biosensors, in cell microarrays and for the fabrication of polymer-based microencapsulators.

Gramse, G., Dols-Perez, A., Edwards, M. A., Fumagalli, L., Gomila, G., (2013). Nanoscale measurement of the dielectric constant of supported lipid bilayers in aqueous solutions with electrostatic force microscopy Biophysical Journal 104, (6), 1257-1262

We present what is, to our knowledge, the first experimental demonstration of dielectric constant measurement and quantification of supported lipid bilayers in electrolyte solutions with nanoscale spatial resolution. The dielectric constant was quantitatively reconstructed with finite element calculations by combining thickness information and local polarization forces which were measured using an electrostatic force microscope adapted to work in a liquid environment. Measurements of submicrometric dipalmitoylphosphatidylcholine lipid bilayer patches gave dielectric constants of εr ∼ 3, which are higher than the values typically reported for the hydrophobic part of lipid membranes (εr ∼ 2) and suggest a large contribution of the polar headgroup region to the dielectric response of the lipid bilayer. This work opens apparently new possibilities in the study of biomembrane electrostatics and other bioelectric phenomena.

Gomila, G., Esteban-Ferrer, D., Fumagalli, L., (2013). Quantification of the dielectric constant of single non-spherical nanoparticles from polarization forces: Eccentricity effects Nanotechnology 24, (50), 505713

We analyze by means of finite-element numerical calculations the polarization force between a sharp conducting tip and a non-spherical uncharged dielectric nanoparticle with the objective of quantifying its dielectric constant from electrostatic force microscopy (EFM) measurements. We show that for an oblate spheroid nanoparticle of given height the strength of the polarization force acting on the tip depends linearly on the eccentricity, e, of the nanoparticle in the small eccentricity and low dielectric constant regimes (1 < e < 2 and 1 < εr; < 10), while for higher eccentricities (e > 2) the dependence is sub-linear and finally becomes independent of e for very large eccentricities (e > 30). These results imply that a precise account of the nanoparticle shape is required to quantify EFM data and obtain the dielectric constants of non-spherical dielectric nanoparticles. Experimental results obtained on polystyrene, silicon dioxide and aluminum oxide nanoparticles and on single viruses are used to illustrate the main findings.

Gramse, G., Edwards, M.A., Fumagalli, L., Gomila, G., (2013). Theory of amplitude modulated electrostatic force microscopy for dielectric measurements in liquids at MHz frequencies Nanotechnology 24, (41), 415709

A theoretical analysis of amplitude modulated electrostatic force microscopy (AM-EFM) in liquid media at MHz frequencies, based on a simple tip–sample parallel plate model, is presented. The model qualitatively explains the main features of AM-EFM in liquid media and provides a simple explanation of how the measured electric forces are affected by: the frequency of the applied voltage, the tip–sample distance, the ionic concentration, the relative dielectric constant of the solution, and the relative dielectric constant and thickness of the sample. These results provide a simple framework for the design of AM-EFM measurements for localized dielectric characterization in liquid media.

Dols-Perez, A., Sisquella, X., Fumagalli, L., Gomila, G., (2013). Optical visualization of ultrathin mica flakes on semitransparent gold substrates Nanoscale Research Letters 8, (1), 1-5

We show that optical visualization of ultrathin mica flakes on metallic substrates is viable using semitransparent gold as substrates. This enables to easily localize mica flakes and rapidly estimate their thickness directly on gold substrates by conventional optical reflection microscopy. We experimentally demonstrate it by comparing optical images with atomic force microscopy images of mica flakes on semitransparent gold. Present results open the possibility for simple and rapid characterization of thin mica flakes as well as other thin sheets directly on metallic substrates.

Keywords: Atomic force, Conductive AFM, Gold substrates, Metallic substrate, Optical image, Optical reflection, Optical visualization, Ultrathin layers, Atomic force microscopy, Geometrical optics, Gold, Mica, Optical microscopy, Substrates

Fumagalli, Laura, Esteban-Ferrer, Daniel, Cuervo, Ana, Carrascosa, Jose L., Gomila, Gabriel, (2012). Label-free identification of single dielectric nanoparticles and viruses with ultraweak polarization forces Nature Materials Nature Publishing Group 11, (9), 743-826

Label-free detection of the material composition of nanoparticles could be enabled by the quantification of the nanoparticles’ inherent dielectric response to an applied electric field. However, the sensitivity of dielectric nanoscale objects to geometric and non-local effects makes the dielectric response extremely weak. Here we show that electrostatic force microscopy with sub-piconewton resolution can resolve the dielectric constants of single dielectric nanoparticles without the need for any reference material, as well as distinguish nanoparticles that have an identical surface but different inner composition. We unambiguously identified unlabelled ~10unm nanoparticles of similar morphology but different low-polarizable materials, and discriminated empty from DNA-containing virus capsids. Our approach should make the in situ characterization of nanoscale dielectrics and biological macromolecules possible.

Keywords: Biological materials, Nanoscale materials, Characterisation and analytical techniques, Computation, modelling and theory

Calò, A., Sanmartí-Espinal, M., Iavicoli, P., Persuy, M. A., Pajot-Augy, E., Gomila, G., Samitier, J., (2012). Diffusion-controlled deposition of natural nanovesicles containing G-protein coupled receptors for biosensing platforms Soft Matter 8, (46), 11632-11643

Natural vesicles produced from genetically engineered cells with tailored membrane receptor composition are promising building blocks for sensing biodevices. This is particularly true for the case of G-protein coupled receptors (GPCRs) present in many sensing processes in cells, whose functionality crucially depends on their lipid environment. However, the controlled production of natural vesicles containing GPCRs and their reproducible deposition on biosensor surfaces are among the outstanding challenges in the road map to realize practical biomolecular devices based on GPCRs. In this work we present the production and characterization of membrane nanovesicles from Saccharomyces cerevisiae containing heterologously expressed olfactory receptors - a member of the family of GPCRs - and study their deposition onto substrates used as biosensor supports. We show by direct observation with Atomic Force Microscopy that nanovesicles deposit and flatten without rupturing on glass substrates following approximately a diffusive law. We show that surface coverages larger than 20-25% of the substrate can be reproducibly achieved under practical nanovesicle concentrations and reasonable time scales, while keeping to the minimum the presence of background residuals coming from the nanovesicles production process. Surface chemistry modification of gold substrates indicates a higher affinity of natural nanovesicles for acid modified surfaces as compared to amino or alcohol modified surfaces. Present results constitute an important step in the practical realization of biosensor devices based on natural nanovesicles integrating G-protein coupled membrane receptors.

Gramse, G., Gomila, G., Fumagalli, L., (2012). Quantifying the dielectric constant of thick insulators by electrostatic force microscopy: effects of the microscopic parts of the probe Nanotechnology 23, (20), 205703

We present a systematic analysis of the effects that the microscopic parts of electrostatic force microscopy probes (the cone and cantilever) have on the electrostatic interaction between the tip apex and thick insulating substrates (thickness>100mum). We discuss how these effects can influence the measurement and quantification of the local dielectric constant of the substrates. We propose and experimentally validate a general methodology that takes into account the influence of the cone and the cantilever, thus enabling us to obtain very accurate values of the dielectric constants of thick insulators.

Keywords: Polarization, Samples

Gramse, G., Edwards, M. A., Fumagalli, L., Gomila, G., (2012). Dynamic electrostatic force microscopy in liquid media Applied Physics Letters 101, (21), 213108

We present the implementation of dynamic electrostatic force microscopy in liquid media. This implementation enables the quantitative imaging of local dielectric properties of materials in electrolyte solutions with nanoscale spatial resolution. Local imaging capabilities are obtained by probing the frequency-dependent and ionic concentration-dependent electrostatic forces at high frequency (>1 MHz), while quantification of the interaction forces is obtained with finite-element numerical calculations. The results presented open a wide range of possibilities in a number of fields where the dielectric properties of materials need to be probed at the nanoscale and in a liquid environment.

Dols-Perez, Aurora, Fumagalli, Laura, Cohen Simonsen, Adam, Gomila, Gabriel, (2011). Ultrathin spin-coated dioleoylphosphatidylcholine lipid layers in dry conditions: A combined atomic force microscopy and nanomechanical study Langmuir 27, (21), 13165-13172

Atomic force microscopy (AFM) has been used to study the structural and mechanical properties of low concentrated spin-coated dioleoylphosphatidylcholine (DOPC) layers in dry environment (RH approximate to 0%) at the nanoscale. It is shown that for concentrations in the 0.1-1 mM range the structure of the DOPC spin-coated samples consists of an homogeneous lipid monolayer similar to 1.3 nm thick covering the whole substrate on top of which lipid bilayer (or multilayer) micro- and nanometric patches and rims are formed. The thickness of the bilayer structures is found to be similar to 4.5 nm (or multiples of this value for multilayer structures), while the lateral dimensions range from micrometers to tens of nanometer depending on the lipid concentration. The force required to break a bilayer (breakthrough force) is found to be similar to 0.24 nN. No dependence of the mechanical values on the lateral dimensions of the bilayer structures is evidenced. Remarkably, the thickness and breakthrough force values of the bilayers measured in dry environment are very similar to values reported in the literature for supported DOPC bilayers in pure water.

Fumagalli, L., Gramse, G., Esteban-Ferrer, D., Edwards, M. A., Gomila, G., (2010). Quantifying the dielectric constant of thick insulators using electrostatic force microscopy Applied Physics Letters 96, (18), 183107

Quantitative measurement of the low-frequency dielectric constants of thick insulators at the nanoscale is demonstrated utilizing ac electrostatic force microscopy combined with finite-element calculations based on a truncated cone with hemispherical apex probe geometry. The method is validated on muscovite mica, borosilicate glass, poly(ethylene naphthalate), and poly(methyl methacrylate). The dielectric constants obtained are essentially given by a nanometric volume located at the dielectric-air interface below the tip, independently of the substrate thickness, provided this is on the hundred micrometer-length scale, or larger.

Keywords: Borosilicate glasses, Finite element analysis, Insulating thin films, Mica, Nanostructured materials, Permittivity, Polymers, Scanning probe microscopy

Toset, J., Gomila, G., (2010). Three-dimensional manipulation of gold nanoparticles with electro-enhanced capillary forces Applied Physics Letters 96, (4), 043117

We demonstrate the possibility to manipulate 25 nm radius gold nanoparticles in the three spatial dimensions with an atomic force microscope with the use of electroenhanced capillary forces. We show that an enhanced water-bridge can be electrostatically induced between a conducting probe and a metallic nanoparticle by the application of a voltage pulse, which is able to exert a pulling capillary force on the nanoparticle strong enough to detach it from the substrate. The nanoparticle can then be moved, attached to the probe, and placed back to the desired location on the substrate simply by contacting it.

Keywords: Atomic force microscopy, Capillarity, Gold, Nanoparticles, Nanotechnology

Sanmarti, M., Iavicoli, P., Pajot-Augy, E., Gomila, G., Samitier, J., (2010). Human olfactory receptors immobilization on a mixed self assembled monolayer for the development of a bioelectronic nose Procedia Engineering (EUROSENSOR XXIV CONFERENCE) 24th Eurosensor Conference (ed. Jakoby, B., Vellekoop, M.J.), Elsevier Science (Linz, Austria) 5, 786-789

The present work focuses on the development of an immunosensing surface to build a portable olfactory system for the detection of complex mixture of odorants. Homogeneous cell derived vesicles expressing the olfactory receptors were produced and immobilized with efficiency onto a gold substrate through an optimized surface functionalization method.

Keywords: Bioelectronic noses, Biosensors, Nanoproteoliposomes, Nanosomes, Olfactory receptors, SAMs

Fumagalli, L., Ferrari, G., Sampietro, M., Gomila, G., (2009). Quantitative nanoscale dielectric microscopy of single-layer supported biomembranes Nano Letters 9, (4), 1604-1608

We present the experimental demonstration of low-frequency dielectric constant imaging of single-layer supported biomembranes at the nanoscale. The dielectric constant image has been quantitatively reconstructed by combining the thickness and local capacitance obtained using a scanning force microscope equipped with a sub-attofarad low-frequency capacitance detector. This work opens new possibilities for studying bioelectric phenomena and the dielectric properties of biological membranes at the nanoscale.

Keywords: Atomic-force microscopy, Nnear-field microscopy, Purple membrane, Scanning capacitance, Biological-systems, Fluid, Spectroscopy, Resolution, Proteins, Dynamics

Gramse, G., Casuso, I., Toset, J., Fumagalli, L., Gomila, G., (2009). Quantitative dielectric constant measurement of thin films by DC electrostatic force microscopy Nanotechnology 20, (39), 395702

A simple method to measure the static dielectric constant of thin films with nanometric spatial resolution is presented. The dielectric constant is extracted from DC electrostatic force measurements with the use of an accurate analytical model. The method is validated here on thin silicon dioxide films (8 nm thick, dielectric constant approximately 4) and purple membrane monolayers (6 nm thick, dielectric constant approximately 2), providing results in excellent agreement with those recently obtained by nanoscale capacitance microscopy using a current-sensing approach. The main advantage of the force detection approach resides in its simplicity and direct application on any commercial atomic force microscope with no need of additional sophisticated electronics, thus being easily available to researchers in materials science, biophysics and semiconductor technology.

Keywords: Roscopy, Membrane, Tip, Polarizability, Polarization, Resolution, Nanotubes, Charge

Rodriguez-Trujillo, R., Castillo-Fernandez, O., Garrido, M., Arundell, M., Valencia, A., Gomila, G., (2008). High-speed particle detection in a micro-Coulter counter with two-dimensional adjustable aperture Biosensors and Bioelectronics 24, (2), 290-296

This article presents the fabrication and characterisation of a high-speed detection micro-Coulter counter with two-dimensional (2D) adjustable aperture and differential impedance detection. The developed device has been fabricated from biocompatible and transparent materials (polymer and glass) and uses the principle of hydrodynamic focusing in two dimensions. The use of a conductive solution for the sample flux and non-conductive solutions for the focalising fluxes provides an adjustable sample flow where particles are aligned and the resistive response concentrated, consequently enhancing the sensitivity and versatility of the device. High-speed counting of 20 mu m polystyrene particles and 5 mu m yeast cells with a rate of up to 1000 particles/s has been demonstrated. Two-dimensional focusing conditions have been used in devices with physical cross-sectional areas of 180 mu m x 65 mu m and 100 mu m x 43 mu m, respectively, in which particles resulted undetectable in the absence of focusing. The 2D-focusing conditions have provided, in addition, increased detection sensitivity by a factor of 1.6 as compared to 1 D-focusing conditions.

Keywords: Impedance, Chip, Microfluidics

Gomila, G., Toset, J., Fumagalli, L., (2008). Nanoscale capacitance microscopy of thin dielectric films Journal of Applied Physics 104, (2), 8

We present an analytical model to interpret nanoscale capacitance microscopy measurements on thin dielectric films. The model displays a logarithmic dependence on the tip-sample distance and on the film thickness-dielectric constant ratio and shows an excellent agreement with finite-element numerical simulations and experimental results on a broad range of values. Based on these results, we discuss the capabilities of nanoscale capacitance microscopy for the quantitative extraction of the dielectric constant and the thickness of thin dielectric films at the nanoscale.

Keywords: AFM, Thickness, Tip

Rodriguez-Trujillo, R., Castillo-Fernandez, O., Arundell, M., Samitier, J., Gomila, G., (2008). Yeast cells detection in a very fast and highly versatile microfabricated cytometer MicroTAS 2008 12th International Conference on Miniaturized Systems for Chemistry and Life Sciences , Chemical and Biological Microsystems Society (San Diego, USA) , 1888-1890

A novel microfluidic chip able to detect a wide range of different cell sizes at very high rates is reported. The device uses two-dimensional hydrodynamic focusing [1] of the sample (conducting) flow by three non-conducting flows and high-speed differential impedance detection electronics. High-speed counting of 15μm polystyrene particles and 5μm yeast cells with a rate of up to 1000 particles/s has been demonstrated. Using of two-dimensional focusing effect turn out to be essential in a device with very large cross-sectional area (100x43 μm2) in which particles result undetectable in the absence of focusing.

Keywords: Coulter-counter, Impedance, Microfluidics, Polydimethylsiloxane

Casuso, I., Pla, M., Gomila, G., Samitier, J., Minic, J., Persuy, M. A., Salesse, R., Pajot-Augy, E., (2008). Immobilization of olfactory receptors onto gold electrodes for electrical biosensor Materials Science & Engineering C 5th Maghreb-Europe Meeting on Materials and their Applicatons for Devices and Physical, Chemical and Biological Sensors , Elsevier Science (Mahdia, TUNISIA) 28, (5-6), 686-691

We investigate the immobilization of native nanovesicles containing functional olfactory receptors onto gold electrodes by means of atomic force microscopy in liquid. We show that nanovesicles can be adsorbed without disrupting them presenting sizes once immobilized ranging from 50 run to 200 nm in diameter. The size of the nanovesicles shows no dependence on the electrode hydrophobicity being constant in a height/width ratio close to 1:3. Nevertheless, electrode hydrophobicity does affect the surface coverage, the surface coverage is five times higher in hydrophilic electrodes than on hydrophobic ones. Surface coverage is also affected by nanovesicles dimensions in suspension, the size homogenization to around 50 nm yields a further five fold increment in surface coverage achieving a coverage of about 50% close to the hard spheres jamming limit (54.7%). A single layer of nanovesicles is always formed with no particle overlap. Present results provide insights into the immobilization on electrodes of olfactory receptors for further olfactory electrical biosensor development.

Keywords: AFM, Adsorption, Odorant, Taste


  • Cypher Atomic Force Microscope (Asylum Research)
  • Nanowizard 4 Bio-Atomic Force Microscope (JPK)
  • Cervantes Atomic Force Microscope (Nanotec Electronica)
  • Easy Scan 2 Atomic Force Microscope (Nanosurf)
  • AxioImager A1m Reflection Optical Microscope (Zeiss) equipped with a AxioCam ERc5s (Zeiss)
  • CompactStat portable electrochemical interface and impedance analyzer (Ivium Technologies)
  • 2 eLockIn204 4-phase Lock-In amplifiers (Anfatec)
  • Keithley 6430 sub-femtoAmp remote sourcemeter (Keithley)


  • Dra. Laura Fumagalli
    University of Manchester, United Kingdom
  • Dr. Ferry Kienberger
    Agilent Technologies Austria, Linz, Austria
  • Prof. Marco Sampietro
    Politecnico di Milano, Italy
  • Dr. Jordi Borrell
    University of Barcelona, Spain
  • Prof. Antonio Juárez
    University of Barcelona, Spain
  • Dr. Manel Puig
    University of Barcelona, Spain

Former members

  • Dr. Laura Fumagalli | Senior Researcher
    Now: Lecturer, School of Physics and Astronomy – Condensed Matter Physics, University of Manchester (UK)

  • Dr. Annalisa Calò | Postdoc
    Now: Postdoc, CUNY Advance Science Research Center (USA)
  • Dr. Aurora Dols-Pérez | Postdoc
    Now: Postdoc at the Technical University of Delft (Nederlands)
  • Dr. Martin Edwards | Postdoc
    Now: Research Assistant Professor, University of Utah (USA)
  • Daniel Esteban Ferrer | PhD Student
  • Georg Gramse | PhD Student
    Now: Senior Researcher, Johannes Kepler University of Linz (Austria)
  • Dr. Jordi Otero | Postdoc
    Now: Postdoc, Institute For Bioengineering of Catalonia (IBEC)
  • Marc van der Hofstadt | PhD Student
  • Maria Chiara Biagi | PhD Student

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