Probing the ionic activation enthalpies in anionic polysaccharide xerogel-based single ion conductor for temperature sensing.

Ionic charge transport in polymer-based solid electrolytes is significantly affected by thermal perturbations, facilitating the detection of temperature variations. However, the impact of ionic interactions and molecular arrangements in polymeric single-ion conductors (SICs) has not been thoroughly investigated for temperature sensing. By probing the effect of the associated energies for ionic interactions and polymeric rearrangements, the thermal sensing characteristics of alginate have been studied. For the first time, alginate SIC interacting with multivalent ions (viz., Na+, Ca2+ and Fe3+) to form xerogel has been exploited as a temperature-sensing layer by fabricating a xerogel-based ionic thermistor (xIT) as a temperature sensor. The xIT has demonstrated stable functioning from 25 to 70 °C and unveiled enhanced sensing abilities in the physiological state of the human body (35-40 °C), exhibiting a monotonic linear response, high sensitivity (-3.77 % °C-1), and high accuracy (0.1 °C). The sensing characteristic is observed due to the inward ionic flux under thermal and electrical perturbations. The concentration of ionic charge carriers and ionic drift are assumed to be Arrhenius-activated processes. A general microscopic model of ion transport within polysaccharides has been elucidated via hopping mechanisms, and the effects of the associated activation energies on temperature sensitivity have been explained.

Soluto-thermal Marangoni convection in stationary micro-bioreactors on heated substrates: Tool for in vitro diagnosis of PSA.

The investigation of antigen-laden droplet deposition patterns on antibody-immobilized substrates has potential for disease detection. Stationary droplets that contain antigens on surfaces immobilized with antibodies can function as microreactors. Temperature modulation enhances reaction efficiency and reduces detection time in droplet-based systems. Thus, the aim of this study is to explore the impact of substrate heating on the structures of protein deposits and the influence of substrate temperature on thermo-solutal Marangoni convection within the droplets. Previous research has explored deposition patterns as diagnostic tools, but limited investigations have focused on the effects of substrate heating on protein deposit structures and the influence of substrate temperature on thermo-solutal Marangoni convection within droplets, creating a knowledge gap. In this study, we conducted experiments to explore how heating the substrate affects the deposition patterns of droplets containing prostate-specific antigen (PSA) on a substrate immobilized with anti-PSA IgG. Additionally, we investigated the thermo-solutal Marangoni convection within these droplets. Our findings reveal distinct deposition patterns classified into dendritic structures (heterogeneous), transitional patterns, and needle-like (homogeneous) structures. The presence of prominent coffee rings and the variation in crystal size across different groups highlight the interplay between thermal and solutal Marangoni advection. Entropy analysis provides insights into structural differences within and between patterns. This work optimizes substrate temperatures for reduced evaporation and detection times while preserving protein integrity, advancing diagnostic tool development, and improving understanding of droplet-based systems.

Highly sensitive Cu-ethylenediamine/PANI composite sensor for NH3 detection at room temperature.

Ammonia detection is needed in several sectors including environmental monitoring, automobile industry, and in medical diagnosis. Conducting polymers, such as polyaniline (PANI), have been utilized to develop NH3 sensors operating at room temperature. However, the performance of these sensors in terms of sensitivity and selectivity need improvement. Functionalization of conducting PANI with metal nanocomposites have shown improved sensor performance. In this work, we report a highly sensitive copper-based nanocomposite for NH3 detection. The novelty lies in utilization of copper-ethylenediamine (Cu-en) nanocomposite functionalized over PANI for gas sensing. Resistance of the 20 wt% Cu-en with PANI increased 3.8 times upon exposure to 100 ppm of NH3. The nanocomposite sensor detected NH3 concentrations as low as 2 ppm. Further, the sensing mechanism was studied by in-situ IV characteristics and impedance spectroscopy during NH3 exposure. NH3 showed ionic interaction with PANI, and Cu2+. The strong affinity of Cu2+ for the lone pair of NH3 enhanced the sensor response from 0.78 to 3.8 for 100 ppm of NH3 at 20 °C. The sensor response was completely recovered after heating at 75 °C, which indicates reusability of the sensor. The sensor showed selectivity for NH3 over ethanol and H2S. The response was reasonably stable after bending the flexible sensor for 1000 times at a radius of 5 mm.

Investigating the Effect of Antibody-Antigen Reactions on the Internal Convection in a Sessile Droplet via Microparticle Image Velocimetry and DLVO Analysis.

The evaporation of antigen-laden sessile droplets on antibody-immobilized PDMS substrates could be used in place of microwells for detection purposes owing to the lesser requirements of analytes and a reduced reaction time. To develop such techniques, the effects of different parameters on the reaction efficiency and on the resulting deposition patterns of antigens on the surface after evaporation need to be well understood. While the resultant deposition patterns from the evaporation of droplets of biological fluids on surfaces are being studied for various biomedical applications, systems where the analyte of interest in the droplet binds to the surface have not been investigated until now. While the effect of temperature on the internal convection within sessile droplets has been studied, the effect of the analyte (antigen in this work) concentration and the analyte-surface (antigen-antibody in this work) binding on the internal convection has not been studied until now. Therefore, to gain insight, the evaporation dynamics of sessile droplets with different concentrations of antigens along with polystyrene microspheres (used as tracers) in phosphate-buffered saline (PBS) on antibody-immobilized PDMS substrates were experimentally studied using microparticle image velocimetry (PIV). It was found that Marangoni flow due to concentration gradients and surface reactions was responsible for the observed velocity field. The antibody-antigen reaction (as compared to the control case of no surface reaction) and higher concentrations of prostate specific antigen (PSA) resulted in increased strength of Marangoni convection. To obtain further insight into the different deposition patterns obtained, the contributions of different particle-particle and particle-substrate forces were determined, and it was observed that the Marangoni forces along with surface tension and DLVO forces create a uniform deposition of the particles present within the droplet. This learning could be used to design biosensors.

pH-Based Detection of Target Analytes in Diluted Serum Samples Using Surface Plasmon Resonance Immunosensor.

Detection of minute quantities of target antigens in serum samples (consisting of a mixture of proteins/biomolecules) can be achieved by enhancement of the capture efficiencies of heterogeneous immunosensors. An important process parameter which affects the capture of target analytes in such immunosensors is the pH of the solution as the target proteins present in the serum samples are charged molecules. Here, we investigated the capture of prostate-specific antigens (PSAs), first in a mixed-analyte system wherein the solution contained two other non-specific proteins along with the target analyte, using the surface plasmon resonance spectroscopy. There are no reports on the detection of antigens in a mixed system based on the optimization of the pH values of the carrier fluid, and this is the motivation of the present work. Further, we studied interference effects caused by the presence of these non-specific proteins in the mixed-analyte systems by artificially increasing the ratio of the interfering proteins to that of the target protein. Eventually PSA spiked into the rabbit serum samples was captured through the optimization of the pH of the solution. We could detect PSA in the serum samples when diluted to 100 times or more, where the amounts of other interfering proteins were ~ 66 times that of the amount of PSA. This study proposes a heterogeneous immunosensor to detect the target analytes in the diluted serum samples by tuning pH the of solution mixture, which can be utilized to detect disease biomarkers in serum samples.

Correlation of Capture Efficiency with the Geometry, Transport, and Reaction Parameters in Heterogeneous Immunosensors.

Higher capture efficiency of biomarkers in heterogeneous immunosensors would enable early detection of diseases. Several strategies are used to improve the capture efficiency of these immunosensors including the geometry of the system along with the transport and reaction parameters. Having a prior knowledge of the behavior of the above parameters would facilitate the design of an efficient immunosensor. While the contributions of the transport and reaction parameters toward understanding of the mechanism involved in capture have been well studied in the literature, their effect in combination with the geometry of the sensors has not been explored until now. In this work, we have experimentally demonstrated that the capture efficiency of the antigen-antibody systems is inversely related to the size of the sensor patch. The experimental system was simulated in order to get an in-depth understanding of the mechanism behind the experimental observation. Further, the extent of heterogeneity in the system was analyzed using the Sips isotherm to obtain the heterogeneity index (α) and the reaction rate constant (K(D)) as fitted parameters for a sensor patch of 1.5 mm radius. The experimental kinetic data obtained for the same sensor patch matched reasonably with the simulation results by considering K(D) as the global affinity constant, which indicated that our system can be considered to be homogeneous. Our simulation results associated with the size dependency of the capture efficiency were in agreement with the trends obtained in our experimental observations where an inverse relation was observed owing to the fact that the mass-transfer limitation decreases with the decrease in the size of the sensor patch. The possible underlying mechanism associated with size dependency of capture efficiency was discussed based on the time-dependent radial variation of captured antigens obtained from our simulation results. A study on the parametric variation was further conducted for the nonmixed and mixed systems on the transport (Deff), reaction (K(D)), and geometric parameters (R). Two different correlations were established for the nonmixed and mixed systems between the capture efficiency (f) and a nondimensional number (t(D)/t(R)) consisting of the above-mentioned parameters. Such unified relations will be useful in designing heterogeneous immunosensors and can be extended to microfluidic immunosensors.

Signal Amplification in Field Effect-Based Sandwich Enzyme-Linked Immunosensing by Tuned Buffer Concentration with Ionic Strength Adjuster.

Miniaturization of the sandwich enzyme-based immunosensor has several advantages but could result in lower signal strength due to lower enzyme loading. Hence, technologies for amplification of the signal are needed. Signal amplification in a field effect-based electrochemical immunosensor utilizing chip-based ELISA is presented in this work. First, the molarities of phosphate buffer saline (PBS) and concentrations of KCl as ionic strength adjuster were optimized to maximize the GOx glucose-based enzymatic reactions in a beaker for signal amplification measured by change in the voltage shift with an EIS device (using 20 µl of solution) and validated with a commercial pH meter (using 3 ml of solution). The PBS molarity of 100 µM with 25 mM KCl provided the maximum voltage shift. These optimized buffer conditions were further verified for GOx immobilized on silicon chips, and similar trends with decreased PBS molarity were obtained; however, the voltage shift values obtained on chip reaction were lower as compared to the reactions occurring in the beaker. The decreased voltage shift with immobilized enzyme on chip could be attributed to the increased Km (Michaelis-Menten constant) values in the immobilized GOx. Finally, a more than sixfold signal enhancement (from 8 to 47 mV) for the chip-based sandwich immunoassay was obtained by altering the PBS molarity from 10 to 100 µM with 25 mM KCl.

Contribution of rotational diffusivity towards the transport of antigens in heterogeneous immunosensors.

Higher capture efficiency in heterogeneous immunosensors is desirable for the detection of cancer biomarkers at low concentrations. The process of the capture of these antigens is transport limited since the rates of antigen/antibody reactions are faster. In the case of non-flow systems, diffusive transport has contributions from both translational and rotational phenomena. Since the contribution of the rotational diffusivity is comparatively less explored in the literature, we have studied the same for three antigens ­ bovine serum albumin (BSA), prostate specific antigen (PSA) and C-reactive proteins (CRP). We quantified the rotational diffusivities using the time resolved fluorescence anisotropy method, and further quantified the contribution of the rotational diffusivities to the overall diffusivity of the antigens, and also studied the effect of the process parameters ­ temperature and pH of the solution. With an increase in temperature, the rotational diffusivity increased showing Arrhenius dependence while with the variation of pH, it showed a non-monotonic behavior having maxima closer to the isoelectric point of the corresponding antigens. This interesting behavior of the pH values could be attributed to lesser electro-viscous effects when the antigen molecule is neutral around its isoelectric point. The optimization of the pH and temperature for the immunosensors could be utilized to design efficient immunosensors.