Optically-induced Dielectrophoresis Research Articles

Optically induced dielectrophoresis for continuous separation and purification of C. vulgaris and H. pluvialis

Marine microalgae are widely present in the natural environment, exhibiting a significant economic value. However, during the inoculation and cultivation process of microalgae, the introduction of unwanted algae is bound to trigger nutrient competition, leading to a decrease in the growth rate of microalgae and consequently impacting their economic value in production. To address this issue, this study integrates the optically induced dielectrophoresis (ODEP) manipulation technology based on the continuous flow in a microfluidic system. A two-stage cell filter, utilizing two virtual optical spots, is designed and manufactured. Leveraging the size differences between microalgae, continuous separation and purification of mixed samples containing Chlorella vulgaris and Haematococcus pluvialis are achieved within microchannels. Additionally, optimal ODEP manipulation conditions for mixed algal liquid samples, comprising C. vulgaris and H. pluvialis, are demonstrated, including appropriate alternating current voltage (6 V), alternating current frequency (100 kHz), light spot width (40 μm), and sample flow rate (0.9 μl/min). Analysis of mixed liquid samples collected at the chip's outlet reveals a reduction in the proportion of H. pluvialis from 37.5% to 1.2% after separation. In summary, this study proposes an ODEP microfluidic system capable of continuously separating and purifying microalgae with different biological characteristics, showcasing its potential as an alternative to traditional labor-intensive microalgae separation techniques.

Nanoscale sorting of extracellular vesicles via optically-induced dielectrophoresis on an integrated microfluidic system.

We reported a microfluidic system for sorting of extracellular vesicles (EVs), which can house DNAs, RNAs, lipids, proteins, and metabolites that are important in intercellular communication. Their presence within bodily fluids has demonstrated potential in both clinical diagnostic and therapeutic applications. Furthermore, EVs exhibit distinct subtypes categorized by their sizes, each endowed with unique biophysical properties. Despite several existing techniques for EV isolation and purification, diminished purity and prolonged processing times still hamper clinical utility; comprehensive capture of EVs remains an ongoing pursuit. To address these challenges, we devised an innovative method for automated sorting of nano-scale EVs employing optically-induced dielectrophoresis on an integrated microfluidic chip. With this approach, EVs of three distinct size categories (small: 100-150 nm, medium-sized: 150-225 nm, and large: 225-350 nm) could be isolated at a purity of 86%. This new method has substantial potential in expediting EV research and diagnostics.

The Utilization of Optically Induced Dielectrophoresis (ODEP)-Based Cell Manipulation in a Microfluidic System for the Purification and Sorting of Circulating Tumor Cells (CTCs) with Different Sizes.

The analysis of circulating tumor cells (CTCs) at the molecular level holds great promise for several clinical applications. For this goal, the harvest of high-purity, size-sorted CTCs with different subtypes from a blood sample are important. For this purpose, a two-step CTC isolation protocol was proposed, by which the immunomagnetic beads-based cell separation was first utilized to remove the majority of blood cells. After that, an optically induced dielectrophoresis (ODEP) microfluidic system was developed to (1) purify the CTCs from the remaining magnetic microbeads-bound blood cells and to (2) sort and separate the CTCs with different sizes. In this study, the ODEP microfluidic system was designed and fabricated. Moreover, its optimum operation conditions and performance were explored. The results exhibited that the presented technique was able to purify and sort the cancer cells with two different sizes from a tested cell suspension in a high-purity (93.5% and 90.1% for the OECM 1 and HA22T cancer cells, respectively) manner. Overall, this study presented a technique for the purification and sorting of cancer cells with different sizes. Apart from this application, the technique is also useful for other applications in which the high-purity and label-free purification and sorting of cells with different sizes is required.

Development of an Optically Induced Dielectrophoresis (ODEP) Microfluidic System for High-Performance Isolation and Purification of Bacteria.

For the rapid detection of bacteria in a blood sample, nucleic acid amplification-based assays are believed to be promising. Nevertheless, the nucleic acids released from the dead blood cells or bacteria could affect the assay performance. This highlights the importance of the isolation of live bacteria from blood samples. To address this issue, this study proposes a two-step process. First, a blood sample was treated with the immuno-magnetic microbeads-based separation to remove the majority of blood cells. Second, an optically induced dielectrophoresis (ODEP) microfluidic system with an integrated dynamic circular light image array was utilized to further isolate and purify the live bacteria from the remaining blood cells based on their size difference. In this work, the ODEP microfluidic system was developed. Its performance for the isolation and purification of bacteria was evaluated. The results revealed that the method was able to harvest the live bacteria in a high purity (90.5~99.2%) manner. Overall, the proposed method was proven to be capable of isolating and purifying high-purity live bacteria without causing damage to the co-existing cells. This technical feature was found to be valuable for the subsequent nucleic-acid-based bacteria detection, in which the interferences caused by the nontarget nucleic acids could be eliminated.

Development of an optically induced dielectrophoresis (ODEP) microfluidic system with a virtual gel filtration chromatography (GFC)-inspired mechanism for the high-performance sorting and separation of microparticles based on their size differences

Size-based sorting and separation of microparticles is critical for many applications. Conventional techniques might not be suitable for this task particularly when sample is limited. Integration of optically induced dielectrophoresis (ODEP) in microfluidic system for the task is believed promising. However, its utilization can be limited by the influence of the friction force acting on microparticles. In this situation, the microparticles might not be effectively sorted and separated. To address this challenge, this study presented an ODEP-based virtual gel filtration chromatography (GFC)-inspired mechanism. By mimicking the concept of GFC, this study proposed to use a circular light image array to maximize the retention time difference among various microparticles when they flowed through such an array. In this study, the array design and its operational conditions were determined. Additionally, its performance for sorting and separation of polystyrene (PS) microbeads with 4 different sizes was evaluated. Results demonstrated that the proposed method was capable of sorting and separating microbeads in a high-performance manner (e.g., purity range: 92–100%). This could be beyond what is currently possible using the current ODEP-based method. Overall, this study presented a new ODEP technique that could solve the problems commonly encountered in ODEP-based microparticle sorting, and separation.

Multi-electrode parallel three-dimensional manipulation based on typical optically-induced dielectrophoresis

In this paper, three-dimensional manipulation based on typical optically-induced dielectrophoresis (ODEP) was realized by using different shapes of electrodes and forced in different directions, and parallel three-dimensional manipulation of multiple electrodes was studied. SMMC-7721 cells were manipulated in 8.5% (w/v) sucrose solution, and the direction of the force on the cell (positive or negative) was determined by the frequency of the external electrical signal. We used light patterns of different shapes (dot, stripe, and ring) to achieve three-dimensional manipulation of cells based on positive force and the constricted ring to achieve three-dimensional manipulation based on negative force. We studied the effect of electrode spacing on manipulation when multiple electrodes were simultaneously used for three-dimensional manipulation. To achieve parallel and independent three-dimensional manipulation of multiple electrodes, it is necessary to ensure a large electrode spacing.

Effects of light size and intensity on photoconductive effect-based optically-induced dielectrophoresis for three-dimensional manipulation

This study aims to study the effect of light size and intensity on photoconductive effect-based optically-induced dielectrophoresis (ODEP) for three-dimensional manipulation of cells. HL-7702 cells and SMMC-7721 cells in sucrose solution were manipulated three-dimensionally by ODEP. We found that the ODEP force generated by the three-dimensional non-uniform electric field will form a three-dimensional space, and the positive ODEP force will attract the cells near the space into it. When the number of cells is large, the three-dimensional stacking phenomenon will occur, that is, the three-dimensional assembly of cells. When the size of the light spot is similar to that of the cell, the projector with weak light intensity can only manipulate a single cell, while the laser with higher light intensity can carry out cell three-dimensional assembly. When the light intensity is similar, the larger size of light can produce larger a three-dimensional assembly space. So the cell three-dimensional manipulation by ODEP is achieved only when the photoconductive effect induced by the light pattern reaches the required intensity. The three-dimensional manipulation of cells by ODEP is from plane to stereo, and its realization is related to the light size and the photoconductive layer resistance determined by light intensity. And the three-dimensional space of assembling cells is related to the light size and intensity, and the larger and stronger the light, the larger the space.

Virtual Filter Membranes in a Microfluidic System for Sorting and Separating Size-Based Micro Polystyrene Beads by Illumination Intensity Design in Optically Induced Dielectrophoresis (ODEP)

In biomedical diagnosis, the efficient separation and purification of specific targets from clinical samples is the desired first step. Herein, the concept of virtual filter membranes based on optically-induced dielectrophoresis (ODEP) manipulation in a microfluidic channel is proposed as a light screening membrane for the separation of polystyrene (PS) microparticles with three different diameters of 15.8, 10.8 and 5.8 µm. The ODEP manipulation velocity of three types of PS microparticles reacted with the color brightness setting was investigated to determine the light intensity to induce an ODEP force higher than the drag force of fluid speed. The color brightness of the light bar in three areas of the light screening membrane was selected as 60%, 70% and 100% to isolate PS microparticles with diameters of 15.8, 10.8 and 5.8 µm, respectively. With a double light bar and a flow rate of 3 µL/min, the recovery rate and isolation purity was improved by 95.1~100% and 94.4~98.6% from the mixture of three types of PS microparticles within 2 min, respectively. This proposed light screening membrane could be a candidate for the separation of small-volume and rare biomedical samples, including circulating tumor cells (CTCs) and bacteria in the blood.

The electrodynamics of rod-like microparticles based on optically induced dielectrophoresis.

Due to its characteristics of noncontact, non-damage, high flux, and easy-to-achieve flexible manipulation, optically induced dielectrophoresis (ODEP) technology has been employed to manipulate microspherical biological particles, including separation, enrichment, capture, arrangement, and fusion. However, in nature, biomolecules are morphologically diverse, and some of them are rodlike. In order to illustrate the electrodynamics of rodlike particles under the action of ODEP, a transient multi-physical field coupling model of ODEP chip under the hypothesis of electrical double layer thin layer was established in this paper. The arbitrary Lagrangian-Eulerian method is used to track single-rod particle in the strong coupled flow field and electric field simultaneously. The influence of several key factors, including the applied alternating current (AC) electric voltage, the width of optical bright area, and the initial position of particle, on the trajectory of particle center was analyzed in positive dielectrophoresis (DEP) action and negative DEP action, respectively. Especially, the planar motion process of rodlike particles was discussed together. The research results reveal the electrodynamics behavior of rodlike particles based on the action of ODEP, which may provide theoretical support for the further design of rodlike biological cells manipulation chip based on AC ODEP technology in the future.

The Utilization of Tunable Transducer Elements Formed by the Manipulation of Magnetic Beads with Different Sizes via Optically Induced Dielectrophoresis (ODEP) for High Signal-to-Noise Ratios (SNRs) and Multiplex Fluorescence-Based Biosensing Applications.

Magnetic beads improve biosensing performance by means of their small volume and controllability by magnetic force. In this study, a new technique composed of optically induced dielectrodphoresis (ODEP) manipulation and image processing was used to enhance the signal-to-noise ratio of the fluorescence for stained magnetic beads. According to natural advantages of size-dependent particle isolation by ODEP manipulation, biomarkers in clinical samples can be easily separated by different sizes of magnetic beads with corresponding captured antibodies, and rapidly distinguished by separated location of immunofluorescence. To verify the feasibility of the concept, magnetic beads with three different diameters, including 21.8, 8.7, and 4.2 μm, were easily separated and collected into specific patterns in the defined target zone treated as three dynamic transducer elements to evaluate fluorescence results. In magnetic beads with diameter of 4.2 μm, the lowest signal-to-noise ratio between stained and nonstained magnetic beads was 3.5. With the help of ODEP accumulation and detection threshold setting of 32, the signal-to-noise ratio was increased to 77.4, which makes this method more reliable. With the further optimization of specific antibodies immobilized on different-size magnetic beads in the future, this platform can be a potential candidate for a high-efficiency sensor array in clinical applications.

Accurate Micromanipulation of Optically Induced Dielectrophoresis Based on a Data-Driven Kinematic Model.

The precise control method plays a crucial role in improving the accuracy and efficiency of the micromanipulation of optically induced dielectrophoresis (ODEP). However, the unmeasurable nature of the ODEP force is a great challenge for the precise automatic manipulation of ODEP. Here, we propose a data-driven kinematic model to build an automatic control system for the precise manipulation of ODEP. The kinematic model is established by collecting the input displacement of the optical pattern and the output displacements of the manipulated object. Then, the control system based on the model was designed, and its feasibility and control precise were validated by numerical simulations and actual experiments on microsphere manipulation. In addition, the applications of ODEP manipulation in two typical scenarios further demonstrated the feasibility of the designed control system. This work proposes a new method to realize the precise manipulation of ODEP technology by establishing a kinematic model and a control system for micromanipulation, and it also provides a general approach for the improvement of the manipulation accuracy of other optoelectronic tweezers.

Recent advances in optically induced di-electrophoresis and its biomedical applications.

The development of the micro/nano science and technology has promoted the evolvement of human civilization tremendously. The advancement of the micro/nano science and technology highly depends on the progress of the micro/nano manipulation techniques, and the micro/nano-scaled manipulation level is the critical sign of the micro/nano science and technology. This review, aimed at the demand and the challenge of the micro/nano material and biomedical fields and related to the scientific issues and implementation techniques of the optically induced di-electrophoresis (ODEP). We explained its working principle, manipulating method, and influencing factors of ODEP force to a certain extent. A number of application fields based-ODEP technology and specific applications so far are summarized and reviewed. Finally, some perspectives are provided on current development trends, future research directions, and challenges of ODEP.

Implementation of flexible virtual microchannels based on optically induced dielectrophoresis

Micro-nano particle manipulation methods in liquid environments have been widely used in the fields such as medicine, biology and material science. Nevertheless, the methods usually rely on pre-prepared physical microfluidic channels. In this work, virtual electrodes based on the optically induced dielectrophoresis (ODEP) method were used as virtual microchannels instead of traditional physical microfluidic channels. Virtual microchannels with different shapes were implemented by the designs of projected light patterns, which made the virtual microchannels have great flexibility and controllability. The theory of ODEP was verified by simulation and analysis of electric field distributions. The relationship between the manipulation force and the alternating current (AC) voltage or the AC frequency exerted on the cells was assessed. The experimental results indicated that the manipulation force was increased with the increase of the AC voltage, and it was reduced with the increase of the AC frequency. Moreover, different virtual microchannels were designed to carry out the transportation, aggregation and sorting of yeast cells and rat basophilic leukemia cells (RBL-2H3 cells) and the survival rate of the cells was evaluated. This work shows that the virtual microchannels can be flexibly realized by ODEP in liquid environments.

Isolation method of Saccharomyces cerevisiae from red blood cells based on the optically induced dielectrophoresis technique for the rapid detection of fungal infections.

Saccharomyces cerevisiae (S. cerevisiae) has been classically used to treat diarrhea and diarrhea-related diseases. However, in the past two decades, fungal infections caused by S. cerevisiae have been increasing among immunocompromised patients, and it takes too long to isolate S. cerevisiae from blood to diagnose it in time. In this paper, a new method for the isolation and selection of S. cerevisiae from red blood cells (RBC) is proposed by designing a microfluidic chip with an optically-induced dielectrophoresis (ODEP) system. It was verified by theory and experiments that the magnitude and direction of the dielectrophoresis force applied on RBCs and S. cerevisiae are different, which determine that the S. cerevisiae can be isolated from RBCs by the ODEP system. By designing the specific light images and the dynamic separation mode, the optimal operating conditions were experimentally achieved for acquiring higher purity of S. cerevisiae. The purity ranges were up to 95.9%-97.3%. This work demonstrates a promising tool for efficient and effective purification of S. cerevisiae from RBCs and provides a novel method of S. cerevisiae isolation for the timely diagnosis of fungal infections.

POMDP-Based Real-Time Path Planning for Manipulation of Multiple Microparticles via Optoelectronic Tweezers.

With high throughput and high flexibility, optoelectronic tweezers (OETs) hold huge potential for massively parallel micromanipulation. However, the trajectory of the virtual electrode has been planned in advance in most synchronous manipulations for multiple targets based on an optically induced dielectrophoresis (ODEP) mechanism, which is insufficient to ensure the stability and efficiency in an environment with potential collision risk. In this paper, a synchronously discretized manipulation method based on a centralized and decoupled path planner is proposed for transporting microparticles of different types with an OET platform. An approach based on the Kuhn-Munkres algorithm is utilized to achieve the goal assignment between target microparticles and goal positions. With the assistance of a visual feedback module, a path planning approach based on the POMDP algorithm dynamically determines the motion strategies of the particle movement to avoid potential collisions. The geometrical parameters of the virtual electrodes are optimized for different types of particles with the goal of maximum transport speed. The experiments of micropatterning with different morphologies and transporting multiple microparticles (e.g., polystyrene microspheres and 3T3 cells) to goal positions are performed. These results demonstrate that the proposed manipulation method based on optoelectronic tweezers is effective for multicell transport and promises to be used in biomedical manipulation tasks with high flexibility and efficiency.

Label-free purification and characterization of optogenetically engineered cells using optically-induced dielectrophoresis.

Optogenetically engineered cell population obtained by heterogeneous gene expression plays a vital role in life science, medicine, and biohybrid robotics, and purification and characterization are essential to enhance its application performance. However, the existing cell purification methods suffer from complex sample preparation or inevitable damage and pollution. The efficient and nondestructive label-free purification and characterization of the optogenetically engineered cells, HEK293-ChR2 cells, is provided here using an optically-induced dielectrophoresis (ODEP)-based approach. The distinctive crossover frequencies of the engineered cells and the unmodified cells enable effective separation due to the opposite DEP forces on them. The ODEP-based approach can greatly improve the purity of the separated cell population and especially, the ratio of the engineered cells in the separated cell population can be enhanced by 275% at a low transfection rate. The size and the membrane capacitance of the separated cell population decreases and increases, respectively, as the ratio of the engineered cells grows in the cell population, indicating that successful expression of ChR2 in a single HEK293 cell makes its size and membrane capacitance smaller and larger, respectively. The results of biohybrid imaging with the optogenetically engineered cells demonstrated that cell purification can improve the imaging quality. This work proves that the separation and purification of engineered cells are of great significance for their application in practice.

Improvement of Background Solution for Optically Induced Dielectrophoresis-Based Cell Manipulation in a Microfluidic System.

Optically induced dielectrophoresis (ODEP) is effective for cell manipulation. However, its utilization has been limited by the requirement of solution with low conductivity. This issue has been ignored in ODEP-relevant studies. To address this issue, this study aims to investigate to what extent the cell viability and performance of ODEP-based cell manipulation are affected by low conductivity conditions. Additionally, this study aims to modify sucrose solutions to reduce the impacts caused by low-conductivity solutions. Results revealed the use of sucrose solution in ODEP operation could significantly reduce the viability of the manipulated cells by 9.1 and 38.5% after 2- and 4-h incubation, respectively. Prolonged operation time (e.g., 4 h) in sucrose solution could lead to significantly inferior performance of cell manipulation, including 47.2% reduction of ODEP manipulation velocity and 44.4% loss of the cells manipulatable by ODEP. The key finding of this study is that the use of bovine serum albumin (BSA)-supplemented sucrose solution (conductivity: 25–50 μS cm−1) might significantly increase the cell viability by 10.9–14.8% compared with that in sucrose solution after 4 h incubation. Moreover, the ODEP manipulation velocity of cells in the BSA-supplemented sucrose solution (conductivity: 25 μS cm−1) was comparable to that in sucrose solution during 4-h incubation. More importantly, compared with sucrose solution, the use of BSA-supplemented sucrose solution (conductivity: 25–50 μS cm−1) contributed high percentage (80.4–93.5%) of the cells manipulatable by ODEP during 4-h incubation. Overall, this study has provided some fundamental information relevant to the improvement of background solutions for ODEP-based cell manipulation.

Label-free rapid isolation of saccharomyces cerevisiae with optically induced dielectrophoresis-based automatic micromanipulation.

Saccharomyces cerevisiae is well-known in the baking and brewing industries and always used for the preparation of probiotics, especially its subtype, Saccharomyces boulardii, to prevent and treat various diarrhea and intestinal diseases. However, case reports on the side effects of a wide range of serious infections for the elderly, immunocompromised and critically ill patients after treatment with the S. cerevisiae have been increasing in recent years. The existing diagnose methods of the invasive S. cerevisiae infections in clinical, especially, the key step of the method-cell isolation, is time-consuming that always miss timey diagnose and early prevention. Here, we propose a new automatic micromanipulation method to label-free rapid isolation of S. cerevisiae based on the optically-induced dielectrophoresis (ODEP) technology, combining with image processing and recognition. S. cerevisiae is firstly identified by the image recognition method and then, automatically captured and moved to the target location by designing optical patterns. The results indicate the method can flexibly and automatically manipulate multiple S. cerevisiae cells simultaneously, such as, arranging S. cerevisiae cells, moving an array of the cells at any directions, aggregating the cells, and separating S. cerevisiae from the solution mixed with impurities. This work represents a step toward the use of automatic micromanipulation of ODEP technology to automatically and rapidly isolate S. cerevisiae for the detection of the invasive S. cerevisiae infections.

Label-Free Rapid Detection of Invasive S. cerevisiae Infections With Optically Induced Dielectrophoresis-Based Micromanipulation and Graphene Transistor

Saccharomyces cerevisiae (S. cerevisiae) is well-known as baker's yeast and brewer's yeast in the food industries and also used as a probiotic in humans. It has been classically considered as a safe non-pathogenic organism, but the number of cases about its implication as an etiologic agent of invasive infections has been increasing in the last two decades. Furthermore, it is difficult to achieve timely diagnosis and early prevention of the related infections in clinical due to long-time being sped in isolation and detection of S. cerevisiae. Here, we propose a novel method of label-free rapid detection of invasive S. cerevisiae infections with a coupled system of optically induced dielectrophoresis (ODEP)-based micromanipulation and graphene transistor. The S. cerevisiae was quickly separated and isolated from its surroundings by ODEP-based micromanipulation, and its optimal operating conditions were experimentally evaluated to quickly collect S. cerevisiae with higher purity (up to 99%) and larger collection rate (more than 65%). The property of S. cerevisiae was detected accurately by the graphene transistor through two measurement modes. And the results validated the graphene transistor obviously distinguish the different concentrations of the S. cerevisiae, characterized by conductance changes or I-V curves of the graphene transistor. This work represents a step toward the use of the coupling of micromanipulation and nanosensor to realize the integrated isolation and detection of S. cerevisiae. Furthermore, it also has potential as a novel approach of isolation and detection of other organisms for the microbiological research and detection of fungal infections.

Rapid isolation method of Saccharomyces cerevisiae based on optically induced dielectrophoresis technique for fungal infection diagnosis.

Saccharomyces cerevisiae(S. cerevisiae) has been classically used as a treatment for diarrhea and diarrhea-related diseases. However, cases of the fungal infections caused by S. cerevisiae have been increasing in the last two decades among immunocompromised patients, while a long time was spent on S. cerevisiae isolation clinically so it was difficult to achieve timely diagnosis the diseases. Here, a novel approach for isolation and selection of S. cerevisiae is proposed by designing a microfluidic chip with an optically induced dielectrophoresis (ODEP) system. S. cerevisiae was isolated from the surroundings by ODEP due to different dielectrophoretic forces. Two special light images were designed and used to block and separate S. cerevisiae, respectively, and several manipulation parameters of ODEP were experimentally optimized to acquire the maximum isolation efficiency of S. cerevisiae. The results on the S. cerevisiae isolation declared that the purity of the S. cerevisiae selected by the method was up to 99.5%±0.05, and the capture efficiency was up to 65.0%±2.5 within 10 min. This work provides a general method to isolate S. cerevisiae as well as other microbial cells with high accuracy and efficiency and paves a road for biological research in which the isolation of high-purity cells is required.