Publications

Abstract Recent advances in optical trapping have opened new opportunities for manipulating micro and nanoparticles, establishing optical tweezers (OT) as a powerful tool for single-cell analysis. Furthermore, intelligent systems have been developed to characterize these particles, as information about their size and composition can be extracted from the scattered radiation signal. In this manuscript, we aim to explore the potential of optical tweezers for the characterization of sub-micron size variations in microparticles. We devised a case study, aiming to assess the limits of the size discrimination ability of an optical tweezer system, using transparent 4.8 µm PMMA particles, functionalized with streptavidin. We focused on the heavily studied streptavidin-biotin system, with streptavidin-functionalized PMMA particles targeting biotinylated bovine serum albumin. This binding process results in an added molecular layer to the particle’s surface, increasing its radius by approximately 7 nm. An automatic OT system was used to trap the particles and acquire their forward-scattered signals. Then, the signals’ frequency components were analyzed using the power spectral density method followed by a dimensionality reduction via the Uniform Manifold Approximation and Projection algorithm. Finally, a Random Forest Classifier achieved a mean accuracy of 94% for the distinction of particles with or without the added molecular layer. Our findings demonstrate the ability of our technique to discriminate between particles that are or are not bound to the biotin protein, by detecting nanoscale changes in the size of the microparticles. This indicates the possibility of coupling shape-changing bioaffinity tools (such as APTMERS, Molecular Imprinted Polymers, or antibodies) with optical trapping systems to enable optical tweezers with analytical capability. © 2024 SPIE.

Abstract Molecularly imprinted polymers (MIPs) are biomimetic materials of great interest in the scientific and industrial fields for the development of innovative sensing strategies. Herein, we proposed a new sensing application by developing an electrochemical sensor using molecular imprinting (MI) technology for recognition of atrial natriuretic peptide (ANP) both as a free molecule in solution and attached to nanoparticle-based drug delivery systems (DDSs), aiming to provide fast and reliable information on the cell uptake of nanoparticles (NPs). As proof of concept, poly(lactic-co-glycolic acid) (PLGA) NPs were synthesized and used as nanocarriers for ischemic heart disease therapy were synthesized and then functionalized with ANP (named here as PLGANPs@ANP). The MIP receptor film was prepared by electrochemical polymerization of dopamine over the working area of a gold screen-printed electrode (AuSPE), using cyclic voltammetry (CV) technique. The construction of the ANP sensor was carefully optimized to enhance its performance, including the film thickness and the procedures for effective template extraction from the MIP matrix. The MIP biosensor presented a linear response against polymeric NPs (PLGA-NPs@ANP) concentration logarithm ranging from 4.0 mu g mL(-1) to 100 mu g mL(-1), with a sensitivity of - 0.0129 mA mL mu g(-1) decade(-1) and an LOD < 4.0 g mL(-1). Furthermore, the developed MIP receptor film was able to discriminate ANP-functionalized nanocarriers from non-functionalized NPs.

Abstract Sensing at the single cell level can provide insights into its dynamics and heterogeneity, yielding information otherwise unattainable with traditional biological methods where average population behavior is observed. In this context, optical tweezers provide the ability to select, separate, manipulate and identify single cells or other types of microparticles, potentially enabling single cell diagnostics. Forward or backscatter analysis of the light interacting with the trapped cells can provide valuable insights on the cell optical, geometrical and mechanical properties. In particular, the combination of tweezers systems with advanced machine learning algorithms can enable single cell identification capabilities. However, typical processing pipelines require a training stage which often struggles when trying to generalize to new sets of data. In this context, fully automated tweezers system can provide mechanisms to obtain much larger datasets with minimum effort form the users, while eliminating procedural variability. In this work, a pipeline for full automation of optical tweezers systems is discussed. A performance comparison between manually operated and fully automated tweezers systems is presented, clearly showing advantages of the latter. A case study demonstrating the ability of the system to discriminate molecular binding events on microparticles is presented.

Abstract Soil contamination with metals is a major threat for the environment and public health since most metals are toxic to humans and to non-human biota, even at low concentrations. Thus, new sustainable remediation approaches are currently needed to immobilize metals in soils to decrease their mobility and bioavailability. In this work, we explore the application of discarded substrates from hydroponic cultivation, namely coconut shell and a mixture of coconut shell and pine bark, for immobilization of metals (Cd, Cr, Ni, Cu, Pb, Hg, Sb and As) in a naturally contaminated soil from a mining region in Portugal. The immobilization capacity of substrates (added to the soil at 5% mass ratio) was assessed both individually and also combined with other traditional agriculture soil additives (limestone and gypsum, at 2% mass ratio) and nanoparticles of zero-valent iron (nZVI) at 1–3% mass ratio. The overall results obtained after a 30-d incubation showed that the discarded substrates are a viable, economic, and environmental-friendly solution for metal remediation in soils, with the capacity of immobilization ranging from 20 to 91% for the metals and metalloids studied. Furthermore, they showed the capacity to reduce the soil toxicity (EC50 ∼ 6000 mg/L) to non-toxic levels (EC50 > 10000 mg/L) to the bacteria Aliivrio fischeri. © 2024 The Authors

Abstract Over the last decades, the interface between two immiscible electrolyte solutions (ITIES) attracted considerable attention of the scientific community due to their vast applications, such as extraction, catalysis, partition studies and sensing. The aim of this Review is to highlight the potential of electrochemistry at the ITIES for analytical purposes, focusing on ITIES-based sensors for detection and quantification of chemically and biologically relevant (bio)molecules. We start by addressing the evolution of ITIES in terms of number of publications over the years along with an overview of their main applications (Chapter 1). Then, we provide a general historical perspective about pioneer voltammetric studies at water/oil systems (Chapter 2). After that, we discuss the most impacting improvements on ITIES sensing systems from both perspectives, set-up design (interface stabilization and miniaturization, selection of the organic solvent, etc.) and optimization of experimental conditions to improve selectivity and sensitivity (Chapter 3). In Chapter 4, we discuss the analytical applications of ITIES for electrochemical sensing of several types of analytes, including drugs, pesticides, proteins, among others. Finally, we highlight the present achievements of ITIES as analytical tool and provide future challenges and perspectives for this technology (Chapter 5).

Abstract This paper describes the development of an optical tweezers system that operates in fully automatic mode. It features image recognition for particle tracking, allowing for the optical trapping and analysis of identified targets. The system can perform analysis of forward scattered light and Raman spectroscopy of the trapped particles, facilitating the automated analysis of a large number of samples without manual intervention. By leveraging combined analytical methods and AI for robust classification, this system contributes to the advancement of automated diagnostic tools. Preliminary results demonstrate the system's effectiveness using different kinds of standard and biofunctionalized PMMA microparticles.

Abstract Citizen science projects have revealed the potential to promote students’ scientific literacy in its affective and cognitive dimensions. However, in chemistry, these projects are still scarce. Thus, starting from the context of the quality of coastal water and its pollution by (micro)plastics, the citizen science project PVC - Perceiving the Value of Chemistry behind water and microplastics was developed, in which chemistry plays a central role. In order to know the contributions of this project, were investigated the attitudes towards chemistry of 574 Portuguese students (442 from the experimental group and 132 from the control group) of the 3rd Cycle of Basic Education and their chemistry learning at the conceptual and procedural level. To this end, an attitude questionnaire, as a pre- and post-test, and a knowledge test were applied. Moreover, field notes were also collected. In the pre-test, positive attitudes were observed in both experimental and control groups. In the post-test, the experimental group showed significantly more positive attitudinal changes than the control group in all the dimensions presented in the questionnaire. The analysis of the results of the knowledge tests also suggests that the PVC project promoted the conceptual learning of chemistry related to the analysis of the physicochemical parameters of water and polymers. At a procedural level, it has become evident the learning of laboratory techniques and the development of skills such as autonomy, collaboration, and critical thinking, as well as the ability to communicate ideas and results promoted by the engagement in PVC project. © 2024 Facultad de Quimica, UNAM. All rights reserved.

Abstract This study elucidates the perceptions of a group of chemistry teaching undergraduates about the mobilization of knowledge related to the seven bases of the Technological Pedagogical Content Knowledge (TPACK) framework during intervention activities in the pandemic. The descriptive research, with a mixed approach and Survey procedure, was based on observation and 29 Likert scale assertions in a self-report questionnaire, where the nominal variables were reorganized into a scale. Statistical analysis via Statistical Package for the Social Sciences software demonstrated data reliability and normality disparity; thus, parametric (two-way ANOVA) and non-parametric (Kruskal-Wallis) tests were performed, considering a significance level of 5% (p < 0.05). In the end, it was observed that among the 29 assertions, 5 exhibited a significant effect of some group (Gender, Age Group, School Where They Work, Class They Attend, Time of Participation) based on the Kruskal-Wallis test, demonstrating rejection of the null hypothesis (p <= 0.05 not equal H0). Furthermore, the findings indicate a strong need for more holistic initial teacher training, where the use of technology becomes an integral part of Content Knowledge.

Abstract An optical fiber pH sensor based on a long-period fiber grating (LPFG) is reported. Two oppositely charged polymers, polyethylenimine (PEI) and polyacrylic acid (PAA), were alternately deposited on the sensing structure through a layer-by-layer (LbL) electrostatic self-assembly technique. Since the polymers are pH sensitive, their refractive index (RI) varies when the pH of the solution changes due to swelling/deswelling phenomena. The fabricated multilayer coating retained a similar property, enabling its use in pH-sensing applications. The pH of the PAA dipping solution was tuned so that a coated LPFG achieved a pH sensitivity of (6.3 +/- 0.2) nm/pH in the 5.92-9.23 pH range. Only two bilayers of PEI/PAA were used as an overlay, which reduces the fabrication time and increases the reproducibility of the sensor, and its reversibility and repeatability were demonstrated by tracking the resonance band position throughout multiple cycles between different pH solutions. With simulation work and experimental results from a low-finesse Fabry-Perot (FP) cavity on a fiber tip, the coating properties were estimated. When saturated at low pH, it has a thickness of 200 nm and 1.53 +/- 0.01 RI, expanding up to 310 nm with a 1.35 +/- 0.01 RI at higher pH values, mostly due to the structural changes in the PAA.

Abstract Surface plasmon resonance (SPR) conventionally occurs at the interface of a thin metallic film and an external dielectric medium in fiber optics through core-guided light. However, this work introduces theoretical and experimental evidence suggesting that the SPR in optical fibers can also be induced through light scattering from Au nanoparticles (NPs) on the thin metallic film, defined as nanoparticle-induced SPR (NPI-SPR). This method adheres to phase-matching conditions between SPR dispersion curves and the wave vectors of scattered light from Au NPs. Experimentally, these conditions are met on an etched optical fiber, enabling direct interaction between light and immobilized Au NPs. Compared to SPR, NPI-SPR exhibits stronger field intensity in the external region and wavelength tuning capabilities (750 to 1250 nm) by varying Au NP diameters (20 to 90 nm). NPI-SPR demonstrates refractive index sensitivities of 4000 to 4416 nm per refractive index unit, nearly double those of typical SPR using the same optical fiber configuration sans Au NPs. Additionally, NPI-SPR fiber configuration has demonstrated its applicability for developing biosensors, achieving a remarkable limit of detection of 0.004 nm for thrombin protein evaluation, a twenty-fold enhancement compared to typical SPR. These findings underscore the intrinsic advantages of NPI-SPR for sensing. Surface plasmon resonance (SPR) typically occurs at the interface of a thin metallic film and a dielectric medium in fiber optics. This work presents evidence of nanoparticle-induced SPR (NPI-SPR) in optical fibers through light scattering from Au nanoparticles on the thin metallic film. NPI-SPR offers stronger field intensity, wavelength tuning, and enhanced refractive index sensitivities, making it advantageous for biosensing applications. image