Optical Thickness Monitoring as a Strategic Element for the Development of SPR Sensing Applications.

The importance of the monitoring of thickness and rate deposition is indispensable for the fabrication of thin film sensors, such as SPR sensors. The sensitivity of SPR responses varies with the thickness of the film, as well as the linear range. Thus, in the present work, we presented an experimental study of the plasmonic response of Cr/Au thin films deposited onto glass slides by evaporation, based on both a rotation and no-rotation system. The results show that the thickness of the gold film varies from 240 to 620 Å, depending on the glass slide position. The SPR response curves obtained experimentally were compared with simulated plasmonic responses and different parameters such as resonance angle, and the depth, slope and half-width of the SPR curve were analysed.

Early Detection of the Fungal Banana Black Sigatoka Pathogen Pseudocercospora fijiensis by an SPR Immunosensor Method.

Black Sigatoka is a disease that occurs in banana plantations worldwide. This disease is caused by the hemibiotrophic fungus Pseudocercospora fijiensis, whose infection results in a significant reduction in both product quality and yield. Therefore, detection and identification in the early stages of this pathogen in plants could help minimize losses, as well as prevent the spread of the disease to neighboring cultures. To achieve this, a highly sensitive SPR immunosensor was developed to detect P. fijiensis in real samples of leaf extracts in early stages of the disease. A polyclonal antibody (anti-HF1), produced against HF1 (cell wall protein of P. fijiensis) was covalently immobilized on a gold-coated chip via a mixed self-assembled monolayer (SAM) of alkanethiols using the EDC/NHS method. The analytical parameters of the biosensor were established, obtaining a limit of detection of 11.7 µg mL-1, a sensitivity of 0.0021 units of reflectance per ng mL-1 and a linear response range for the antigen from 39.1 to 122 µg mL-1. No matrix effects were observed during the measurements of real leaf banana extracts by the immunosensor. To the best of our knowledge, this is the first research into the development of an SPR biosensor for the detection of P. fijiensis, which demonstrates its potential as an alternative analytical tool for in-field monitoring of black Sigatoka disease.

Enhanced Enzymatic Activity of Laccase (from Pycnoporus sanguineus CS43) Immobilized on Sputtered Nanostructured Gold Thin Films.

Functionalization of thin films with organic ligands has been the subject of intense research due to their potential application as heterogeneous molecular nanosystems. In this work, self-assembled monolayers of thiols (16-mercaptohexadecanoic acid and 11-mercaptoundecanol) were used to bind laccase (from Pycnoporus sanguineus CS43) to nanostructured gold thin films obtained by DC sputtering. Sputtering power, sputtering pressure and substrate temperature were optimized to enhance the activity of the immobilized biomolecules. Scanning electron microscopy, confocal microscopy, X-ray diffraction and UV-vis spectroscopy were used to characterize the SAM-functionalized gold substrates. Our results demonstrate that the highest immobilized enzyme activity values can be achieved on substrates of surface roughness ˜200 nm and Au particle size of about 14 nm. The outstanding quality of the as-prepared substrates makes them particularly attractive as bionanosensors.

Progress in Plasmonic Sensors as Monitoring Tools for Aquaculture Quality Control.

Aquaculture is an expanding economic sector that nourishes the world's growing population due to its nutritional significance over the years as a source of high-quality proteins. However, it has faced severe challenges due to significant cases of environmental pollution, pathogen outbreaks, and the lack of traceability that guarantees the quality assurance of its products. Such context has prompted many researchers to work on the development of novel, affordable, and reliable technologies, many based on nanophotonic sensing methodologies. These emerging technologies, such as surface plasmon resonance (SPR), localised SPR (LSPR), and fibre-optic SPR (FO-SPR) systems, overcome many of the drawbacks of conventional analytical tools in terms of portability, reagent and solvent use, and the simplicity of sample pre-treatments, which would benefit a more sustainable and profitable aquaculture. To highlight the current progress made in these technologies that would allow them to be transferred for implementation in the field, along with the lag with respect to the most cutting-edge plasmonic sensing, this review provides a variety of information on recent advances in these emerging methodologies that can be used to comprehensively monitor the various operations involving the different commercial stages of farmed aquaculture. For example, to detect environmental hazards, track fish health through biochemical indicators, and monitor disease and biosecurity of fish meat products. Furthermore, it highlights the critical issues associated with these technologies, how to integrate them into farming facilities, and the challenges and prospects of developing plasmonic-based sensors for aquaculture.

Novel Thermotolerant Amylase from Bacillus licheniformis Strain LB04: Purification, Characterization and Agar-Agarose.

This study analyzed the thermostability and effect of calcium ions on the enzymatic activity of α-amylase produced by Bacillus licheniformis strain LB04 isolated from Espinazo Hot springs in Nuevo Leon, Mexico. The enzyme was immobilized by entrapment on agar-agarose beads, with an entrapment yield of 19.9%. The identification of the bacteria was carried out using 16s rDNA sequencing. The enzyme was purified through ion exchange chromatography (IEX) in a DEAE-Sephadex column, revealing a protein with a molecular weight of ≈130 kDa. The enzyme was stable at pH 3.0 and heat stable up to 80 °C. However, the optimum conditions were reached at 65 °C and pH 3.0, with a specific activity of 1851.7 U mg-1 ± 1.3. The agar-agarose immobilized α-amylase had a hydrolytic activity nearly 25% higher when compared to the free enzyme. This study provides critical information for the understanding of the enzymatic profile of B. licheniformis strain LB04 and the potential application of the microorganisms at an industrial level, specifically in the food industry.

The Potential Use of a Thin Film Gold Electrode Modified with Laccases for the Electrochemical Detection of Pyrethroid Metabolite 3-Phenoxybenzaldehyde.

There is increasing interest in developing portable technologies to detect human health threats through hybrid materials that integrate specific bioreceptors. This work proposes an electrochemical approach for detecting 3-Phenoxybenzaldehyde (3-PBD), a biomarker for monitoring human exposure to pyrethroid pesticides. The biosensor uses laccase enzymes as an alternative recognition element by direct oxidation of 3-PBD catalysts by the enzyme onto thin-film gold electrodes. The thin-film gold electrode modified by the immobilized laccase was characterized by Fourier-transform infrared spectrometry and scanning electron microscopy. The detection method's electrochemical parameters were established, obtaining a linear range of 5 t 50 µM, the limit of detection, and quantification of 0.061 and 2.02 µM, respectively. The proposed biosensor's analytical performance meets the concentration of pyrethroids detected in natural environments, reflecting its potential as an alternative analytical tool for monitoring the pyrethroid insecticide's presence.

A Novel Enzyme-Based SPR Strategy for Detection of the Antimicrobial Agent Chlorophene.

Chlorophene is an important antimicrobial agent present in disinfectant products which has been related to health and environmental effects, and its detection has been limited to chromatographic techniques. Thus, there is a lack of research that attempts to develop new analytical tools, such as biosensors, that address the detection of this emerging pollutant. Therefore, a new biosensor for the direct detection of chlorophene in real water is presented, based on surface plasmon resonance (SPR) and using a laccase enzyme as a recognition element. The biosensor chip was obtained by covalent immobilization of the laccase on a gold-coated surface through carbodiimide esters. The analytical parameters accomplished resulted in a limit of detection and quantification of 0.33 mg/L and 1.10 mg/L, respectively, fulfilling the concentrations that have already been detected in environmental samples. During the natural river's measurements, no significant matrix effects were observed, obtaining a recovery percentage of 109.21% ± 7.08, which suggested that the method was suitable for the fast and straightforward analysis of this contaminant. Finally, the SPR measurements were validated with an HPLC method, which demonstrated no significant difference in terms of precision and accuracy, leading to the conclusion that the biosensor reflects its potential as an alternative analytical tool for the monitoring of chlorophene in aquatic environments.

Characterization of interdigitated electrode structures for water contaminant detection using a hybrid voltage divider and a vector network analyzer.

Interdigitated capacitive electrode structures have been used to monitor or actuate over organic and electrochemical media in efforts to characterize biochemical properties. This article describes a method to perform a pre-characterization of interdigitated electrode structures using two methods: a hybrid voltage divider (HVD) and a vector network analyzer (VNA). Both methodologies develop some tests under two different conditions: free air and bi-distilled water media. Also, the HVD methodology is used for other two conditions: phosphate buffer with laccase (polyphenoloxidase; EC 1.10.3.2) and contaminated media composed by a mix of phosphate buffer and 3-ethylbenzothiazoline-6-sulfonic acid (ABTS). The purpose of this study is to develop and validate a characterization methodology using both, a hybrid voltage divider and VNA T-# network impedance models of the interdigitated capacitive electrode structure that will provide a shunt RC network of particular interest in detecting the amount of contamination existing in the water solution for the media conditions. This methodology should provide us with the best possible sensitivity in monitoring water contaminant media characteristics. The results show that both methods, the hybrid voltage divider and the VNA methodology, are feasible in determining impedance modeling parameters. These parameters can be used to develop electric interrogation procedures and devices such as dielectric characteristics to identify contaminant substances in water solutions.