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1.
Chemosphere ; 363: 142873, 2024 Sep.
Article in English | MEDLINE | ID: mdl-39019187

ABSTRACT

Electro-osmosis has been valued as a promising technology to enhance the dewatering of waste sludge, stabilization and environmental remediation of soils with low permeability. However, the coefficient of electro-osmotic permeability (keo) is commonly taken as constant value which is particularly not the case in variable charge soil. As a result, the nonlinearity of the electro-osmotic flow (EOF) and the direction reverse could not be interpreted. Herein, the electro-chemical parameters were monitored in electro-osmotic experiment with natural variable charge soil. It was observed that the evolutions showed significant nonlinear behavior and were correlated. The comprehensive Zeta potential model proposed by the authors was applied to simulate the nonlinear keo induced by the variable pH and electrolyte concentration. The agreement between tested and simulated flow rate variation and excess pore water pressure distribution demonstrated the reliability of the theory. The error rate of the simulations through coupling nonlinear keo and voltage gradient Ex was reduced to 29.4% from 381.9% of calculations with constant parameters. The direction reverse of EOF was innovatively interpreted. Hence, the numerical model would act as a useful tool to connect these electro-chemical parameters and provide guidance to evaluate contributions of commonly used pH conditioning measurements.


Subject(s)
Osmosis , Soil , Soil/chemistry , Hydrogen-Ion Concentration , Electroosmosis/methods , Permeability , Models, Theoretical , Environmental Restoration and Remediation/methods , Electrolytes/chemistry
2.
Chemosphere ; 362: 142661, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38906191

ABSTRACT

Electro-osmosis offers an effective method for dewatering and remediating low permeability soil. Long-term observations on nonlinear behavior of electro-osmosis and the influencing factors are not commonly reported. Connection between cessation and direction reversal of electro-osmotic flow (EOF), and the evolution of electro-chemical parameters inside of the soil mass thus remains unclear. The dynamic response of EOF in variable charge soil could be significant, whereas the investigations on which are currently lacking. A series of electro-osmotic experiments were performed with two natural variable charge soils. The results indicated that initial electro-osmotic rate was positively proportional to electric current and initial electrical conductivity of the pore fluid, which could be explained by the ion migration model. The dynamic evolution of electro-osmotic rate and electro-chemical parameters corresponding to the solute and pH conditionings at the electrode compartments demonstrated that: 1) coupling effects of non-uniform distribution of voltage gradient and pH determined the magnitude and direction of EOF rate; 2) compared to the final pHIEP value, the bigger, close and smaller values of the novel index "voltage gradient weighed mean of spatial pH″ represented the forward, terminated and reversed EOF respectively; 3) the classical Helmholtz-Smoluchowski model are proved to be more applicable interpreting the coupled nonlinearity of electro-osmosis during the later steady phase. This work would facilitate future research for a comprehensive electro-osmotic model, and provide guidance to condition the initial and boundary conditions in application of electro-osmotic dewatering and electrokinetic remediation.


Subject(s)
Electric Conductivity , Electroosmosis , Soil , Soil/chemistry , Electroosmosis/methods , Osmosis , Hydrogen-Ion Concentration
3.
PLoS One ; 19(4): e0302150, 2024.
Article in English | MEDLINE | ID: mdl-38625994

ABSTRACT

Electroosmosis has been proposed as a technique to reduce moisture and thus increase the stability of soft clay. However, its high energy consumption and uneven reinforcement effect has limited its popularization and application in practical engineering. This paper presents the results of some electrokinetic tests performed on clayey specimens with different electrification time and anode boundary conditions. The results indicate that the timing of the formation of electroosmotic flow (EF) by the water originally contained in different soil cross sections, from the anode to the cathode, varies. The measuring soil cross section nearest the anode first reached the limiting water content of 22%±3% and electroosmosis had to be stopped. Water injection into the anode during electroosmosis enhanced further drainage of other four measuring soil cross sections until the second soil cross section from the anode reached the limiting water content of 30%±2%. Electroosmosis with water injection into the anode technique provides more uniform reinforcement, increasing EF, and environmental protection. The experimental results highlighted the relevant and expected contribution of water injection into the anode on the effectiveness of the electroosmotic treatment as a soft clay improvement technique.


Subject(s)
Electroosmosis , Soil Pollutants , Clay , Electroosmosis/methods , Soil Pollutants/analysis , Soil , Water
4.
Langmuir ; 40(18): 9622-9629, 2024 May 07.
Article in English | MEDLINE | ID: mdl-38652583

ABSTRACT

The detection and identification of nanoscale molecules are crucial, but traditional technology comes with a high cost and requires skilled operators. Solid-state nanopores are new powerful tools for discerning the three-dimensional shape and size of molecules, enabling the translation of molecular structural information into electric signals. Here, DNA molecules with different shapes were designed to explore the effects of electroosmotic forces (EOF), electrophoretic forces (EPF), and volume exclusion on electric signals within solid-state nanopores. Our results revealed that the electroosmotic force was the main driving force for single-stranded DNA (ssDNA), whereas double-stranded DNA (dsDNA) was primarily dominated by electrophoretic forces in nanopores. Moreover, dsDNA caused greater amplitude signals and moved faster through the nanopore due to its larger diameter and carrying more charges. Furthermore, at the same charge level and amount of bases, circular dsDNA exhibited a tighter structure compared to brush DNA, resulting in a shorter length. Consequently, circular dsDNA caused higher current-blocking amplitudes and faster passage speeds. The characterization approach based on nanopores allows researchers to get molecular information about size and shape in real time. These findings suggest that nanopore detection has the potential to streamline nanoscale characterization and analysis, potentially reducing both the cost and complexity.


Subject(s)
DNA , Nanopores , DNA/chemistry , Nucleic Acid Conformation , DNA, Single-Stranded/chemistry , Electroosmosis/methods
5.
Electrophoresis ; 45(11-12): 1054-1064, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38506142

ABSTRACT

Numerical modeling of Taylor dispersion analysis (TDA) was performed using COMSOL Multiphysics to facilitate better and faster optimization of the experimental conditions. Parameters, such as pressure, electric field, diameter, and length of capillary on the TDA conditions, were examined for particles with hydrodynamic radius (Rh) of 2.5-250 Å. The simulations were conducted using 25, 50, and 100 cm length tubes with diameters of 25, 50, and 100 µm. It was shown that particles with larger diffusion coefficients gave more accurate results at higher velocities, and in longer and wider columns; particles with smaller diffusion coefficients gave more accurate results at smaller velocities, and in shorter and thinner columns. Moreover, the effect of electric field on the validity and the applicability of TDA was studied using TDA in conjunction with capillary electrophoresis. Diffusion coefficients were obtained using a pressure and the TDA equation and compared with those obtained with a pressure in combination of an electric field for fluorescein, FD4, FD20, FD70, and FD500. We found that TDA can be used with the presence of moderate electrophoretic migration and electroosmotic flow, when appropriate conditions were met.


Subject(s)
Electrophoresis, Capillary , Electrophoresis, Capillary/methods , Computer Simulation , Diffusion , Models, Theoretical , Electroosmosis/methods , Electricity , Hydrodynamics , Particle Size , Pressure
6.
Biosensors (Basel) ; 14(3)2024 Feb 20.
Article in English | MEDLINE | ID: mdl-38534219

ABSTRACT

The rotation of cells is of significant importance in various applications including bioimaging, biophysical analysis and microsurgery. Current methods usually require complicated fabrication processes. Herein, we proposed an induced charged electroosmosis (ICEO) based on a chip manipulation method for rotating cells. Under an AC electric field, symmetric ICEO flow microvortexes formed above the electrode surface can be used to trap and rotate cells. We have discussed the impact of ICEO and dielectrophoresis (DEP) under the experimental conditions. The capabilities of our method have been tested by investigating the precise rotation of yeast cells and K562 cells in a controllable manner. By adjusting the position of cells, the rotation direction can be changed based on the asymmetric ICEO microvortexes via applying a gate voltage to the gate electrode. Additionally, by applying a pulsed signal instead of a continuous signal, we can also precisely and flexibly rotate cells in a stepwise way. Our ICEO-based rotational manipulation method is an easy to use, biocompatible and low-cost technique, allowing rotation regardless of optical, magnetic or acoustic properties of the sample.


Subject(s)
Electricity , Electroosmosis , Acoustics , Electrodes , Electroosmosis/methods , Rotation , Humans
7.
Electrophoresis ; 45(13-14): 1265-1266, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38376122

ABSTRACT

A serious error exists in the paper: Alharbi KAM, Riaz A, Sikandar S. An entropy model for Carreau nanofluid ciliary flow with electroosmosis and thermal radiations: a numerical study. Electrophoresis. 2024;45:1112-1129.


Subject(s)
Electroosmosis , Entropy , Electroosmosis/methods , Models, Theoretical , Electrophoresis/methods
8.
Electrophoresis ; 45(15-16): 1307-1315, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38342679

ABSTRACT

Cationic surfactant coatings (e.g., CTAB) are commonly used in CE to control EOF and thereby improve separation efficiencies. However, our understanding of surfactant adsorption and desorption dynamics under EOF conditions is limited. Here, we apply automated zeta potential analysis to study the adsorption and desorption kinetics of CTAB in a capillary under different transport conditions: diameter, length, voltage alternation pattern and frequency, and applied pressure. In contrast to other studies, we observe slower kinetics at distinct capillary wall zeta potential ranges. Within these ranges, which we call "stagnant regimes," the EOF mobility significantly counteracts the electrophoretic (EP) mobility of CTA+ and hinders the net transport. By constructing a numerical model to compare with our experiments and recasting our experimental data in terms of the net CTA+ transport volume normalized by surface area, we reveal that the EP mobility of CTA+ and the capillary surface-area-to-volume ratio dictate the zeta potential range and the duration of the stagnant regime and thereby govern the overall reaction kinetics. Our results indicate that further transport-oriented studies can significantly aid in the understanding and design of electrokinetic systems utilizing CTAB and other charged surfactants.


Subject(s)
Cetrimonium Compounds , Cetrimonium , Electrophoresis, Capillary , Cetrimonium/chemistry , Adsorption , Electrophoresis, Capillary/methods , Cetrimonium Compounds/chemistry , Kinetics , Surface-Active Agents/chemistry , Models, Chemical , Electroosmosis/methods
9.
Electrophoresis ; 45(7-8): 676-686, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38350722

ABSTRACT

Understanding electrokinetic transport in nanochannels and nanopores is essential for emerging biological and electrochemical applications. The viscoelectric effect is an important mechanism implicated in the increase of local viscosity due to the polarization of a solvent under a strong electric field. However, most analyses of the viscoelectric effect have been limited to numerical analyses. In this work, we present a set of analytical solutions applicable to the physical description of viscoelectric effects in nanochannel electrokinetic systems. To achieve such closed-form solutions, we employ the Debye-Hückel approximation of small diffuse charge layer potentials compared to the thermal potential. We analyze critical parameters, including electroosmotic flow profiles, electroosmotic mobility, flow rate, and channel conductance. We compare and benchmark our analytical solutions with published predictions from numerical models. Importantly, we leverage these analytical solutions to identify essential thermophysical and nondimensional parameters that govern the behavior of these systems. We identify scaling parameters and relations among surface charge density, ionic strength, and nanochannel height.


Subject(s)
Electroosmosis , Electroosmosis/methods , Viscosity , Nanotechnology/methods , Microfluidic Analytical Techniques/instrumentation , Microfluidic Analytical Techniques/methods , Nanopores , Osmolar Concentration , Nanostructures/chemistry
10.
Electrophoresis ; 45(13-14): 1155-1170, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38115169

ABSTRACT

Drug delivery systems, where the nanofluid flow with electroosmosis and mixed convection can help in efficient and targeted drug delivery to specific cells or organs, could benefit from understanding the behavior of nanofluids in biological systems. In current work, authors have studied the theoretical model of two-dimensional ciliary flow of blood-based (Eyring-Powell) nanofluid model with the insertion of ternary hybrid nanoparticles along with the effects of electroosmosis, magnetohydrodynamics, thermal radiations, and mixed convection. Moreover, the features of entropy generation are also taken into consideration. The system is modeled in a wave frame with the approximations of large wave number and neglecting turbulence effects. The problem is solved numerically by using the shooting method with the assistance of computational software "Mathematica" for solving the governing equation. According to the temperature curves, the temperature will increase as the Hartman number, fluid factor, ohmic heating, and cilia length increase. It is also disclosed that ternary hybrid nanoparticles result in a change in flow rate when other problem parameters are varied, and the same is true for temperature graphs. Engineers and scientists can make better use of nanofluid-based cooling systems in electronics, automobiles, and industrial processes with the aid of the study's findings.


Subject(s)
Convection , Electroosmosis , Entropy , Electroosmosis/methods , Nanoparticles/chemistry , Models, Theoretical , Nanotechnology/instrumentation , Nanotechnology/methods , Hydrodynamics , Drug Delivery Systems/instrumentation
11.
Anal Bioanal Chem ; 415(20): 4861-4873, 2023 Aug.
Article in English | MEDLINE | ID: mdl-37382654

ABSTRACT

Determining the physical and chemical properties of biologically important particles such as cells, organelles, viruses, exosomes, complexes, nucleotides, and proteins is needed to understand their function. These properties are determined with common analytical tools (mass spectrometry, cryo-EM, NMR, various spectroscopies, nucleotide sequencing, etc.) whose function can be improved when samples are pure and concentrated. Separations science plays a central role in conditioning samples, ranging from low-resolution benchtop operations like precipitations or extractions to higher-resolution chromatography and electrophoresis. In the last two decades, gradient insulator-based dielectrophoresis (g-iDEP) has emerged as a high-resolution separation technique capable of highly selective enrichment of cells, viruses, exosomes, and proteins. Specific evidence has been shown that pure homogeneous and concentrated fractions of cells and exosomes can be generated from complex mixtures. However, recovering those fractions for analysis has not been developed, limiting the technique to an analytical rather than a preparative one. Here, a finite element analysis was undertaken to identify geometries and operational parameters to efficiently remove the enriched fraction while retaining maximum concentration and providing total mass transfer. Geometric factors (e.g., side channel width and distance from the gradient-inducing gap) were studied, along with the addition of a second inlet side channel. Two flow-generating mechanisms-electroosmosis and hydrostatic pressure-were evaluated for semi-optimized device designs, including a comparison of the one- and two-inlet designs. Simulations indicate effectively one hundred percent mass transfer and a concentration increase by an order of magnitude for several device configurations and operational parameters.


Subject(s)
Electroosmosis , Microfluidic Analytical Techniques , Electrophoresis/methods , Electroosmosis/methods , Lab-On-A-Chip Devices
12.
Electrophoresis ; 44(5-6): 558-562, 2023 03.
Article in English | MEDLINE | ID: mdl-36495094

ABSTRACT

Electroosmotic flow (EOF) was determined in tridimensional (3D)-printed microchannels with dimensions smaller than 100 µm. Fused deposition modeling 3D printing using thermoplastic filaments of PETG (polyethylene terephthalate glycol), PLA (polylactic acid), and ABS (acrylonitrile butadiene styrene) were used to fabricate the microchannels. The current monitoring method and sodium phosphate solutions at different pH values (3-10) were used for the EOF mobility (µEOF ) measurements, which ranged from 2.00 × 10-4 to 12.52 × 10-4  cm2  V-1  s-1 . The highest and the smallest µEOF were obtained for the PLA and PETG microchannels, respectively. Adding the cationic surfactant cetyltrimethylammonium bromide to the sodium phosphate solution caused EOF direction reversion in all the studied microchannels. The obtained results can be interesting for developing 3D-printed microfluidic devices, in which EOF is relevant.


Subject(s)
Electroosmosis , Phosphates , Electroosmosis/methods , Printing, Three-Dimensional
13.
Electrophoresis ; 44(1-2): 44-52, 2023 01.
Article in English | MEDLINE | ID: mdl-35775948

ABSTRACT

We have investigated the role of viscoelectric effect on diffusioosmotic flow (DOF) through a nanochannel connected with two reservoirs. The transport equations governing the flow dynamics are solved numerically using the finite element technique. We have extensively analyzed the variation of induced field due to electric double layer (EDL) phenomenon, relative viscosity as modulated by the viscoelectric effect as well as reservoir's concentration difference, and their eventual impact on the underlying flow characteristics. It is revealed that the induced electric field in the EDL enhances fluid viscosity substantially near the charged wall at a higher concentration. We have shown that neglecting viscoelectric effect in the paradigm of diffusioosmotic transport overestimates the net throughput, particularly at a higher concentration difference. Furthermore, we show that pertaining to chemiosmosis dominated regime, the average flow velocity modifies with the increase in concentration difference up to a critical value. In comparison, the rise in the strength of resistive electroosmotic actuation by the accumulation of anions in the upstream reservoir reduces the average flow velocity at a higher concentration difference. We have reported a reduction in critical concentration with the increase in viscoelectric effect. The inferences of this analysis are deemed pertinent to reveal the bearing of viscoelectric effect as a flow control mechanism pertaining to DOF at nanoscale.


Subject(s)
Electricity , Electroosmosis , Electroosmosis/methods
14.
Biosensors (Basel) ; 12(10)2022 Sep 23.
Article in English | MEDLINE | ID: mdl-36290922

ABSTRACT

The performance of receptor-based biosensors is often limited by either diffusion of the analyte causing unreasonable long assay times or a lack of specificity limiting the sensitivity due to the noise of nonspecific binding. Alternating current (AC) electrokinetics and its effect on biosensing is an increasing field of research dedicated to address this issue and can improve mass transfer of the analyte by electrothermal effects, electroosmosis, or dielectrophoresis (DEP). Accordingly, several works have shown improved sensitivity and lowered assay times by order of magnitude thanks to the improved mass transfer with these techniques. To realize high sensitivity in real samples with realistic sample matrix avoiding nonspecific binding is critical and the improved mass transfer should ideally be specific to the target analyte. In this paper we cover recent approaches to combine biosensors with DEP, which is the AC kinetic approach with the highest selectivity. We conclude that while associated with many challenges, for several applications the approach could be beneficial, especially if more work is dedicated to minimizing nonspecific bindings, for which DEP offers interesting perspectives.


Subject(s)
Biosensing Techniques , Reaction Time , Biosensing Techniques/methods , Electroosmosis/methods , Diffusion , Electrophoresis/methods
15.
Small Methods ; 6(8): e2200318, 2022 08.
Article in English | MEDLINE | ID: mdl-35656741

ABSTRACT

During the past decades, scientists have developed different kinds of nanorobots based on various driving principles to realize controlled manipulation of them for potential applications like medical diagnosis and directed cargo delivery. In order to design a nanorobot with advantages of simple operation and precise control that can enrich the family of intelligent nanorobots, an encoding manipulation method is proposed to control the movement of a DNA-nanoparticle assembled nanorobot by combing electrophoresis and electroosmosis effect in independently charged array nanopores. The nanorobot is composed of one nanoparticle and one or two ssDNAs. ssDNAs act as the legs of the nanorobot. The selective ion transport through charged nanopores can induce cooperation and competition between the electroosmosis and electrophoresis, which is the main power to activate the nanorobot. Thus by simply switching the applied electric field and surface charge density of each nanopore which is defined as the encoded nanopore according to a predetermined strategy, the well-controlled encoding manipulation including capturing, releasing, jumping, and crawling of the nanorobot is realized in this work. The study is expected to realize its value in many interesting applications like drug delivery, nanosurgery, and so on in the near future.


Subject(s)
Nanoparticles , Nanopores , DNA , Drug Delivery Systems , Electroosmosis/methods
16.
Electrophoresis ; 43(16-17): 1755-1764, 2022 09.
Article in English | MEDLINE | ID: mdl-35736538

ABSTRACT

This study presents on-chip continuous accumulation and concentration of nanoscale samples using a cascade alternating current electroosmosis (cACEO) flow. ACEO can generate flow motion caused by ion movement due to interactions between the AC electric field and the induced charge layer on the electrode surface, with the potential to accumulate particles, especially in low-conductive liquid. However, the intrinsic particle diffusive motion, which is sensitive to particle size, is an essential element influencing accumulation efficiency. In this study, an electrode combining chevron and double-gap geometry embedded in a microfluidic channel was developed to perform efficient three-dimensional (3D) nanoparticle focusing using ACEO. The chevron electrode pattern was introduced upstream of the focusing zone to overcome particle accumulation in scattering zones near the channel sidewall. To demonstrate the efficiency of the proposed device for particle accumulation, three nanoparticle types were used: latex, metal, and biomaterial. Continuous 3D concentration of 50-nm polystyrene particles was confirmed. The concentration factor, determined based on image processing, became quite high when 50-nm gold nanoparticles were used. Moreover, nanoparticles with a 20-nm diameter were accumulated using cACEO. Finally, we used the concentrator chip to accumulate 50-nm liposome particles, confirming that the device could also successfully concentrate biomaterials.


Subject(s)
Metal Nanoparticles , Microfluidic Analytical Techniques , Electrodes , Electroosmosis/methods , Gold
17.
ACS Nano ; 16(5): 7701-7712, 2022 05 24.
Article in English | MEDLINE | ID: mdl-35435659

ABSTRACT

The voltage-dependent transport through biological and artificial nanopores is being used in many applications such as DNA or protein sequencing and sensing. The primary approach to determine the transport has been to measure the temporal ion current fluctuations caused by solutes when applying external voltages. Crossing the nanoscale confinement in the presence of an applied electric field primarily relies on two factors, i.e., the electrophoretic drag and electroosmosis. The electroosmotic flow (EOF) is a voltage-dependent ion-associated flow of solvent molecules, i.e., usually water, and depends on many factors, such as pH, temperature, pore diameter, and also the concentration of ions. The exact interplay between these factors is so far poorly understood. In this joint experimental and computational study, we have investigated the dependence of the EOF on the concentration of the buffer salt by probing the transport of α-cyclodextrin molecules through the ΔCymA channel. For five different KCl concentrations in the range between 0.125 and 2 M, we performed applied-field molecular dynamics simulations and analyzed the ionic flow and the EOF across the ΔCymA pore. To our surprise, the concentration-dependent net ionic flux changes non-monotonically and nonlinearly and the EOF is seen to follow the same pattern. On the basis of these findings, we were able to correlate the concentration-dependent EOF with experimental kinetic constants for the translocation of α-cyclodextrin through the ΔCymA nanopore. Overall, the results further improve our understanding of the EOF-mediated transport through nanopores and show that the EOF needs to seriously be taken into consideration when analyzing the permeation of (neutral) substrates through nanopores.


Subject(s)
Nanopores , alpha-Cyclodextrins , Electroosmosis/methods , DNA/chemistry , Electrophoresis , Ions
18.
Anal Methods ; 14(18): 1782-1787, 2022 05 13.
Article in English | MEDLINE | ID: mdl-35475508

ABSTRACT

Pressure-assisted electrokinetic injection (PAEKI) was applied for stacking of positively charged biogenic amines (BAs) to improve the sensitivity of capillary electrophoresis (CE). It is well known that the essential step for PAEKI is finding a stationary state of the running buffer such that the movement of the running buffer due to electroosmotic flow (EOF) is counterbalanced by external pressure in the opposite direction of the EOF under a given electric field. In order to find the balance point systematically and integrally, we studied the velocity of the whole BGE in the capillary by the impetus of opposite direction pressure (-0.1 to -0.6 psi), and the velocity of EOF with different voltages. According to the two sets of linear data, the EOF of CE coupled with PAEKI could be counterbalanced at the opposite direction pressure (-0.1 psi) and voltage (7.8 kV). In this study, the injection time was extended up to 0.35 min for all BAs and 0.70 min for the direct ultraviolet (UV) detection of BAs. Compared with hydrodynamic injection (HDI), the enrichment factors for sample injection times of 0.35 min and 0.70 min were 480-fold and 970-fold, respectively. The limits of detection (LODs) (S/N = 3) of indirect and direct UV detection were respectively 8.7-24.3 nmol L-1 and 0.4-4.5 nmol L-1, which reaches the sensitivity of high-performance liquid chromatography-mass spectrophotometry (HPLC-MS). With appropriate sample dilution, PAEKI can be used in the analysis of BAs in chicken.


Subject(s)
Electroosmosis , Electrophoresis, Capillary , Biogenic Amines , Electroosmosis/methods , Electrophoresis, Capillary/methods , Limit of Detection
19.
J Am Chem Soc ; 144(7): 3063-3073, 2022 02 23.
Article in English | MEDLINE | ID: mdl-35143193

ABSTRACT

Protein sequencing, as well as protein fingerprinting, has gained tremendous attention in the electrical sensing realm of solid-state nanopores and is challenging due to fast translocations and the use of high molar electrolytes. Despite providing an appreciable signal-to-noise ratio, high electrolyte concentrations can have adverse effects on the native protein structure. Herein, we present a thorough investigation of low electrolyte sensing conditions across a broad pH and voltage range generating conductive pulses (CPs) irrespective of protein net charge. We used Cas9 as the model protein and demonstrated that unfolding is noncooperative, represented by the gradual elongation or stretching of the protein, and sensitive to both the applied voltage and pH (i.e., charge state). The magnitude of unfolding and the isoelectric point (pI) of Cas9 was found to be correlated and a critical factor in our experiments. Electroosmotic flow (EOF) was always aligned with the transit direction, whereas electrophoretic force (EPF) was either reinforcing (pH < pI) or opposing (pH > pI) the protein's movement, which led to slower translocations at higher pH values. Further exploration of higher pH values led to slowing down of protein with > 30% of the population being slower than 0.5 ms. Our results would be critical for protein sensing at very low electrolytes and to retard their translocation speed without resorting to high-bandwidth equipment.


Subject(s)
CRISPR-Associated Protein 9/metabolism , Nanopores , Electroosmosis/instrumentation , Electroosmosis/methods , Hydrogen-Ion Concentration , Isoelectric Point , Protein Conformation , Protein Unfolding
20.
Electrophoresis ; 43(12): 1259-1262, 2022 06.
Article in English | MEDLINE | ID: mdl-34755360

ABSTRACT

We describe an improved method for determining the electroosmotic mobility and zeta potential of surfaces based on a current-monitoring method. This technique eliminates the requirement for measurements of channel dimensions and sample conductivities, leading to a simple high precision measurement. The zeta potential of PDMS is measured for native surfaces and surfaces treated with a nonionic surfactant in low-conductivity electrolytes.


Subject(s)
Microfluidic Analytical Techniques , Microfluidics , Electrolytes , Electroosmosis/methods , Microfluidic Analytical Techniques/methods
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