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Dhiman, Shikha, Andrian, Teodora, Gonzalez, Beatriz Santiago, Tholen, Marrit ME., Wang, Yuyang, Albertazzi, Lorenzo, (2022). Can super-resolution microscopy become a standard characterization technique for materials chemistry? Chemical Science 13, 2152-2166

The characterization of newly synthesized materials is a cornerstone of all chemistry and nanotechnology laboratories. For this purpose, a wide array of analytical techniques have been standardized and are used routinely by laboratories across the globe. With these methods we can understand the structure, dynamics and function of novel molecular architectures and their relations with the desired performance, guiding the development of the next generation of materials. Moreover, one of the challenges in materials chemistry is the lack of reproducibility due to improper publishing of the sample preparation protocol. In this context, the recent adoption of the reporting standard MIRIBEL (Minimum Information Reporting in Bio–Nano Experimental Literature) for material characterization and details of experimental protocols aims to provide complete, reproducible and reliable sample preparation for the scientific community. Thus, MIRIBEL should be immediately adopted in publications by scientific journals to overcome this challenge. Besides current standard spectroscopy and microscopy techniques, there is a constant development of novel technologies that aim to help chemists unveil the structure of complex materials. Among them super-resolution microscopy (SRM), an optical technique that bypasses the diffraction limit of light, has facilitated the study of synthetic materials with multicolor ability and minimal invasiveness at nanometric resolution. Although still in its infancy, the potential of SRM to unveil the structure, dynamics and function of complex synthetic architectures has been highlighted in pioneering reports during the last few years. Currently, SRM is a sophisticated technique with many challenges in sample preparation, data analysis, environmental control and automation, and moreover the instrumentation is still expensive. Therefore, SRM is currently limited to expert users and is not implemented in characterization routines. This perspective discusses the potential of SRM to transition from a niche technique to a standard routine method for material characterization. We propose a roadmap for the necessary developments required for this purpose based on a collaborative effort from scientists and engineers across disciplines.

Keywords: blinking, fluorophore, intramolecular spirocyclization, localization, nanoparticles, resolution limit, reveals, single-molecule fluorescence, stimulated-emission, Characterization techniques, Diffraction, Distributed computer systems, Environmental management, Information reporting, Material chemistry, Materials characterization, Minimum information, Optical reconstruction microscopy, Optical resolving power, Sample preparation, Structure dynamics, Structure functions, Super-resolution microscopy, Synthesized materials


Arista-Romero M, Delcanale P, Pujals S, Albertazzi L, (2022). Nanoscale Mapping of Recombinant Viral Proteins: From Cells to Virus-Like Particles ACS Photonics 9, 101-109

Influenza recombinant proteins and virus-like particles (VLPs) play an important role in vaccine development (e.g., CadiFluS). However, their production from mammalian cells suffers from low yields and lack of control of the final VLPs. To improve these issues, characterization techniques able to visualize and quantify the different steps of the process are needed. Fluorescence microscopy represents a powerful tool able to image multiple protein targets; however, its limited resolution hinders the study of viral constructs. Here, we propose the use of super-resolution microscopy and in particular of DNA-point accumulation for imaging in nanoscale topography (DNA-PAINT) microscopy as a characterization method for recombinant viral proteins on both cells and VLPs. We were able to quantify the amount of the three main influenza proteins (hemagglutinin (HA), neuraminidase (NA), and ion channel matrix protein 2 (M2)) per cell and per VLP with nanometer resolution and single-molecule sensitivity, proving that DNA-PAINT is a powerful technique to characterize recombinant viral constructs.

Keywords: dna-paint, hemagglutinin, influenza, neuraminidase, paint, recombinant proteins, single-molecule localization microscopy, single-particle analysis, virus-like particles, Dna-paint, Hemagglutinin, Influenza, Neuraminidase, Paint, Recombinant proteins, Single particle analysis, Single-molecule localization microscopy, Single-particle analysis, Super-resolution microscopy, Superresolution microscopy, Virus-like particles


Sans J, Arnau M, Estrany F, Turon P, Alemán C, (2021). Regulating the Superficial Vacancies and OH− Orientations on Polarized Hydroxyapatite Electrocatalysts Advanced Materials Interfaces 8

Smart designs of hydroxyapatite (HAp) materials with customized electrical properties are drawing increasing attention for their wide range of potential applications. Such enhanced electrical properties directly arise from the number and orientation of OH groups in the HAp lattice. Although different polarization treatments have been proposed to enhance the final conductivity by generating vacancies at high temperatures and imposing specific OH orientations through electric voltages, no direct measurement showing the evolution that OH groups undergo has been described yet. In this article, the first direct empirical observation that allows the characterization of both the generation of vacancies and the polarization of OH groups is reported. The mechanisms behind the electrical enhancement are elucidated allowing to distinguish between charge accumulation at the crystal grains, which is due to the formed vacancies, and charge accumulation in the boundaries of particles. In addition, a linear dependence between the number of vacancies and the superficial charge is observed. Therefore, it is demonstrated that the charge accumulation at the micrometric grain boundaries has a great impact on the catalytic properties of the thermally stimulated polarized HAp. These results will be used for further optimization of the catalyst properties. − − − −

Keywords: electrocatalysts, hydroxyl orientation, thermally stimulated polarization, vacancies, Charge delocalization, Electrocatalysts, Hydroxyl orientation, Thermally stimulated polarization, Vacancies


Andrian T, Bakkum T, van Elsland DM, Bos E, Koster AJ, Albertazzi L, van Kasteren SI, Pujals S, (2021). Super-resolution correlative light-electron microscopy using a click-chemistry approach for studying intracellular trafficking Methods in Cell Biology 162, 303-331

© 2020 Elsevier Inc. Correlative light and electron microscopy (CLEM) entails a group of multimodal imaging techniques that are combined to pinpoint to the location of fluorescently labeled molecules in the context of their ultrastructural cellular environment. Here we describe a detailed workflow for STORM-CLEM, in which STochastic Optical Reconstruction Microscopy (STORM), an optical super-resolution technique, is correlated with transmission electron microscopy (TEM). This protocol has the advantage that both imaging modalities have resolution at the nanoscale, bringing higher synergies on the information obtained. The sample is prepared according to the Tokuyasu method followed by click-chemistry labeling and STORM imaging. Then, after heavy metal staining, electron microscopy imaging is performed followed by correlation of the two images. The case study presented here is on intracellular pathogens, but the protocol is versatile and could potentially be applied to many types of samples.

Keywords: cells, click-chemistry, complex, correlative light and electron microscopy, cycloaddition, ligation, localization, proteins, resolution limit, single molecule localization microscopy, stochastic optical reconstruction microscopy (storm), storm, super-resolution microscopy, tokuyasu cryo-sectioning, tool, Click-chemistry, Correlative light and electron microscopy, Fluorescent-probes, Single molecule localization microscopy, Stochastic optical reconstruction microscopy (storm), Super-resolution microscopy, Tokuyasu cryo-sectioning, Transmission electron microscopy


Delcanale, P., Albertazzi, L., (2020). DNA-PAINT super-resolution imaging data of surface exposed active sites on particles Data in Brief 30, 105468

Surface functionalization with targeting ligands confers to nanomaterials the ability of selectively recognize a biological target. Therefore, a quantitative characterization of surface functional molecules is critical for the rational development of nanomaterials-based applications, especially in nanomedicine research. Single-molecule localization microscopy can provide visualization of surface molecules at the level of individual particles, preserving the integrity of the material and overcoming the limitations of analytical methods based on ensemble averaging. Here we provide single-molecule localization data obtained on streptavidin-coated polystyrene particles, which can be exploited as a model system for surface-functionalized materials. After loading of the active sites of streptavidin molecules with a biotin-conjugated probe, they were imaged with a DNA-PAINT imaging approach, which can provide single-molecule imaging at subdiffraction resolution and molecule counting. Both raw records and analysed data, consisting in a list of space-time single-molecule coordinates, are shared. Additionally, Matlab functions are provided that analyse the single-molecule coordinates in order to quantify features of individual particles. These data might constitute a valuable reference for applications of similar quantitative imaging methodologies to other types of functionalized nanomaterials.

Keywords: DNA-PAINT, Functional materials, Nanoparticles, Single-molecule localization microscopy, Super-resolution microscopy


Burgués, Javier, Marco, Santiago, (2020). Feature extraction for transient chemical sensor signals in response to turbulent plumes: Application to chemical source distance prediction Sensors and Actuators B: Chemical 320, 128235

This paper describes the design of a linear phase low-pass differentiator filter with a finite impulse response (FIR) for extracting transient features of gas sensor signals (the so-called “bouts”). The detection of these bouts is relevant for estimating the distance of a gas source in a turbulent plume. Our current proposal addresses the shortcomings of previous ‘bout’ estimation methods, namely: (i) they were based in non-causal digital filters precluding real time operation, (ii) they used non-linear phase filters leading to waveform distortions and (iii) the smoothing action was achieved by two filters in cascade, precluding an easy tuning of filter performance. The presented method is based on a low-pass FIR differentiator, plus proper post-processing, allowing easy algorithmic implementation for real-time robotic exploration. Linear phase filters preserve signal waveform in the bandpass region for maximum reliability concerning both ‘bout’ detection and amplitude estimation. As a case study, we apply the proposed filter to predict the source distance from recordings obtained with metal oxide (MOX) gas sensors in a wind tunnel. We first perform a joint optimization of the cut-off frequency of the filter and the bout amplitude threshold, for different wind speeds, uncovering interesting relationships between these two parameters. We demonstrate that certain combinations of parameters can reduce the prediction error to 8 cm (in a distance range of 1.45 m) improving previously reported performances in the same dataset by a factor of 2.5. These results are benchmarked against traditional source distance estimators such as the mean, variance and maximum of the response. We also study how the length of the measurement window affects the performance of different signal features, and how to select the filter parameters to make the predictive models more robust to changes in wind speed. Finally, we provide a MATLAB implementation of the bout detection algorithm and all analysis code used in this study.

Keywords: Gas sensors, Differentiator, Low pass filter, Metal oxide semiconductor, MOX sensors, Signal processing, Feature extraction, Gas source localization, Robotics


Burgués, Javier, Hernández, Victor, Lilienthal, Achim J., Marco, Santiago, (2020). Gas distribution mapping and source localization using a 3D grid of metal oxide semiconductor sensors Sensors and Actuators B: Chemical 304, 127309

The difficulty to obtain ground truth (i.e. empirical evidence) about how a gas disperses in an environment is one of the major hurdles in the field of mobile robotic olfaction (MRO), impairing our ability to develop efficient gas source localization strategies and to validate gas distribution maps produced by autonomous mobile robots. Previous ground truth measurements of gas dispersion have been mostly based on expensive tracer optical methods or 2D chemical sensor grids deployed only at ground level. With the ever-increasing trend towards gas-sensitive aerial robots, 3D measurements of gas dispersion become necessary to characterize the environment these platforms can explore. This paper presents ten different experiments performed with a 3D grid of 27 metal oxide semiconductor (MOX) sensors to visualize the temporal evolution of gas distribution produced by an evaporating ethanol source placed at different locations in an office room, including variations in height, release rate and air flow. We also studied which features of the MOX sensor signals are optimal for predicting the source location, considering different lengths of the measurement window. We found strongly time-varying and counter-intuitive gas distribution patterns that disprove some assumptions commonly held in the MRO field, such as that heavy gases disperse along ground level. Correspondingly, ground-level gas distributions were rarely useful for localizing the gas source and elevated measurements were much more informative. We make the dataset and the code publicly available to enable the community to develop, validate, and compare new approaches related to gas sensing in complex environments.

Keywords: Mobile robotic olfaction, Metal oxide gas sensors, Signal processing, Sensor networks, Gas source localization, Gas distribution mapping


Burgués, Javier, Hernández, Victor, Lilienthal, Achim J., Marco, Santiago, (2019). Smelling nano aerial vehicle for gas source localization and mapping Sensors 19, (3), 478

This paper describes the development and validation of the currently smallest aerial platform with olfaction capabilities. The developed Smelling Nano Aerial Vehicle (SNAV) is based on a lightweight commercial nano-quadcopter (27 g) equipped with a custom gas sensing board that can host up to two in situ metal oxide semiconductor (MOX) gas sensors. Due to its small form-factor, the SNAV is not a hazard for humans, enabling its use in public areas or inside buildings. It can autonomously carry out gas sensing missions of hazardous environments inaccessible to terrestrial robots and bigger drones, for example searching for victims and hazardous gas leaks inside pockets that form within the wreckage of collapsed buildings in the aftermath of an earthquake or explosion. The first contribution of this work is assessing the impact of the nano-propellers on the MOX sensor signals at different distances to a gas source. A second contribution is adapting the ‘bout’ detection algorithm, proposed by Schmuker et al. (2016) to extract specific features from the derivative of the MOX sensor response, for real-time operation. The third and main contribution is the experimental validation of the SNAV for gas source localization (GSL) and mapping in a large indoor environment (160 m2) with a gas source placed in challenging positions for the drone, for example hidden in the ceiling of the room or inside a power outlet box. Two GSL strategies are compared, one based on the instantaneous gas sensor response and the other one based on the bout frequency. From the measurements collected (in motion) along a predefined sweeping path we built (in less than 3 min) a 3D map of the gas distribution and identified the most likely source location. Using the bout frequency yielded on average a higher localization accuracy than using the instantaneous gas sensor response (1.38 m versus 2.05 m error), however accurate tuning of an additional parameter (the noise threshold) is required in the former case. The main conclusion of this paper is that a nano-drone has the potential to perform gas sensing tasks in complex environments.

Keywords: Robotics, Signal processing, Electronics, Gas source localization, Gas distribution mapping, Gas sensors, Drone, UAV, MOX sensor, Quadcopter


Burgues, J., Marco, S., (2019). Feature extraction of gas sensor signals for gas source localization ISOEN 2019 18th International Symposium on Olfaction and Electronic Nose , IEEE (Fukuoka, Japan) , 1-3

This paper explores which signal features of a gas sensor are optimum for assessing the proximity to a gas source in an open environment. Specifically, we compare three statistical descriptors of the signal (mean, variance and maximum response) against the 'bout' frequency, a feature computed in the derivative of the response. The experimental setup includes a generator of turbulent plumes and a sensing board composed of three metal oxide (MOX) sensors of different types. The main conclusion is that the maximum response is the most robust feature across the three sensors. The 'bout' frequency can be very sensitive to an additional parameter (the noise threshold).

Keywords: Feature extraction, Gas plume, Gas sensors, Gas source localization, MOX, Signal processing


Burgués, Javier, Hernandez, Victor, Lilienthal, Achim J., Marco, Santiago, (2018). 3D Gas distribution with and without artificial airflow: An experimental study with a grid of metal oxide semiconductor gas sensors Proceedings EUROSENSORS 2018 , MDPI (Graz, Austria) 2, (13), 911

Gas distribution modelling can provide potentially life-saving information when assessing the hazards of gaseous emissions and for localization of explosives, toxic or flammable chemicals. In this work, we deployed a three-dimensional (3D) grid of metal oxide semiconductor (MOX) gas sensors deployed in an office room, which allows for novel insights about the complex patterns of indoor gas dispersal. 12 independent experiments were carried out to better understand dispersion patters of a single gas source placed at different locations of the room, including variations in height, release rate and air flow profiles. This dataset is denser and richer than what is currently available, i.e., 2D datasets in wind tunnels. We make it publicly available to enable the community to develop, validate, and compare new approaches related to gas sensing in complex environments.

Keywords: MOX, Metal oxide, Flow visualization, Gas sensors, Gas distribution mapping, Sensor grid, 3D, Gas source localization, Indoor


Khalil, I. S. M., Magdanz, V., Sánchez, S., Schmidt, O. G., Misra, S., (2015). Precise localization and control of catalytic janus micromotors using weak magnetic fields International Journal of Advanced Robotic Systems , 12, (2), 1-7

We experimentally demonstrate the precise localization of spherical Pt-Silica Janus micromotors (diameter 5 μm) under the influence of controlled magnetic fields. First, we control the motion of the Janus micromotors in two-dimensional (2D) space. The control system achieves precise localization within an average region-of-convergence of 7 μm. Second, we show that these micromotors provide sufficient propulsion force, allowing them to overcome drag and gravitational forces and move both downwards and upwards. This propulsion is studied by moving the micromotors in three-dimensional (3D) space. The micromotors move downwards and upwards at average speeds of 19.1 μm/s and 9.8 μm/s, respectively. Moreover, our closed-loop control system achieves localization in 3D space within an average region-of-convergence of 6.3 μm in diameter. The precise motion control and localization of the Janus micromotors in 2D and 3D spaces provides broad possibilities for nanotechnology applications.

Keywords: 3D space, Localization, Magnetic control, Micromotors, Self-propulsion


Palleja, T., Balsa, R., Tresanchez, M., Moreno, J., Teixido, M., Font, D., Marco, S., Pomareda, V., Palacin, J., (2014). Corridor gas-leak localization using a mobile Robot with a photo ionization detector sensor Sensor Letters , 12, (6-7), 974-977

The use of an autonomous mobile robot to locate gas-leaks and air quality monitoring in indoor environments are promising tasks that will avoid risky human operations. However, these are challenging tasks due to the chaotic gas profile propagation originated by uncontrolled air flows. This paper proposes the localization of an acetone gas-leak in a 44 m-length indoor corridor with a mobile robot equipped with a PID sensor. This paper assesses the influence of the mobile robot velocity and the relative height of the PID sensor in the profile of the measurements. The results show weak influence of the robot velocity and strong influence of the relative height of the PID sensor. An estimate of the gas-leak location is also performed by computing the center of mass of the highest gas concentrations.

Keywords: Gas source detection, LIDAR sensor, Mobile robot, PID sensor, SLAM, Acetone, Air quality, Gases, Indoor air pollution, Mobile robots, Robots, Air quality monitoring, Autonomous Mobile Robot, Gas sources, Indoor environment, Leak localization, LIDAR sensors, Profile propagation, SLAM, Ionization of gases


Martínez, Dani, Pallejà, T., Moreno, Javier, Tresanchez, Marcel, Teixidó, M., Font, Davinia, Pardo, Antonio, Marco, Santiago, Palacín, Jordi, (2014). A mobile robot agent for gas leak source detection Advances in Intelligent Systems and Computing Trends in Practical Applications of Heterogeneous Multi-Agent Systems. The PAAMS Collection (ed. Bajo Perez, Javier, Corchado Rodríguez, Juan M., Mathieu, Philippe, Campbell, Andrew, Ortega, Alfonso, Adam, Emmanuel, Navarro, Elena M., Ahrndt, Sebastian, Moreno, Maríaa N., Julián, Vicente), Springer International Publishing 293, 19-25

This paper presents an autonomous agent for gas leak source detection. The main objective of the robot is to estimate the localization of the gas leak source in an indoor environment without any human intervention. The agent implements an SLAM procedure to scan and map the indoor area. The mobile robot samples gas concentrations with a gas and a wind sensor in order to estimate the source of the gas leak. The mobile robot agent will use the information obtained from the onboard sensors in order to define an efficient scanning path. This paper describes the measurement results obtained in a long corridor with a gas leak source placed close to a wall.

Keywords: Gas detection, Mobile robot agent, Laser sensor, Self-localization


Hernando, Jordi, Hoogenboom, Jacob, van Dijk, Erik, Garcia-Parajo, Maria, van Hulst, Niek F., (2008). Ultrafast single-molecule photonics: Excited state dynamics in coherently coupled complexes Journal of Luminescence 16th International Conference on Dynamical Processes in Excited States of Solids (ed. -----), Elsevier Science BV (Segovia, Spain) 128, (5-6), 1050-1052

We present a single-molecule study on femtosecond dynamics in multichromophoric systems, combining fs pump-probe, emission-spectra and fluorescence-lifetime analysis. The ultrafast fs approach gives direct information on the initial exciton dynamics after excitation. The lifetime data show superradiance, a direct measure for the extent of the coherent coupling and static disorder. The spectra finally reveal the role of exciton-phonon coupling. At the single-molecule level a wide range of exciton delocalization lengths and energy redistribution times is revealed.

Keywords: Single-molecule detection, Pump-probe, Exciton delocalization, Superradiance, Exciton-phonon coupling