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1.
Sci Rep ; 14(1): 13293, 2024 06 10.
Artigo em Inglês | MEDLINE | ID: mdl-38858424

RESUMO

We introduce magnetophoresis-based microfluidics for sorting biological targets using positive Magnetophoresis (pM) for magnetically labeled particles and negative Magnetophoresis (nM) for label-free particles. A single, externally magnetized ferromagnetic wire induces repulsive forces and is positioned across the focused sample flow near the main channel's closed end. We analyze magnetic attributes and separation performance under two transverse dual-mode magnetic configurations, examining magnetic fields, hydrodynamics, and forces on microparticles of varying sizes and properties. In pM, the dual-magnet arrangement (DMA) for sorting three distinct particles shows higher magnetic gradient generation and throughput than the single-magnet arrangement (SMA). In nM, the numerical results for SMA sorting of red blood cells (RBCs), white blood cells (WBCs), and prostate cancer cells (PC3-9) demonstrate superior magnetic properties and throughput compared to DMA. Magnetized wire linear movement is a key design parameter, allowing device customization. An automated device for handling more targets can be created by manipulating magnetophoretic repulsion forces. The transverse wire and magnet arrangement accommodate increased channel depth without sacrificing efficiency, yielding higher throughput than other devices. Experimental validation using soft lithography and 3D printing confirms successful sorting and separation, aligning well with numerical results. This demonstrates the successful sorting and separating of injected particles within a hydrodynamically focused sample in all systems. Both numerical and experimental findings indicate a separation accuracy of 100% across various Reynolds numbers. The primary channel dimensions measure 100 µm in height and 200 µm in width. N52 permanent magnets were employed in both numerical simulations and experiments. For numerical simulations, a remanent flux density of 1.48 T was utilized. In the experimental setup, magnets measuring 0.5 × 0.5 × 0.125 inches and 0.5 × 0.5 × 1 inch were employed. The experimental data confirm the device's capability to achieve 100% separation accuracy at a Reynolds number of 3. However, this study did not explore the potential impact of increased flow rates on separation accuracy.


Assuntos
Técnicas Analíticas Microfluídicas , Humanos , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/métodos , Separação Celular/métodos , Separação Celular/instrumentação , Eritrócitos , Microfluídica/métodos , Microfluídica/instrumentação , Leucócitos , Hidrodinâmica , Linhagem Celular Tumoral
2.
Lab Chip ; 23(18): 4117-4125, 2023 09 13.
Artigo em Inglês | MEDLINE | ID: mdl-37655531

RESUMO

The lysis of cancer cells inside a sessile droplet was performed using traveling surface acoustic waves (SAWs) without any chemical reagents. Raman spectrum profiling was then carried out to explore detailed cell-derived data. The Rayleigh waves formed by an interdigital transducer were made to propagate along the surface of an LiNbO3 substrate. Polystyrene microparticles (PSMPs) were used to establish mechanical cell lysis effectively, and gold nanoparticles (AuNPs) were added to enhance the Raman signals from the lysed cells by SAWs. The lysis efficiency was evaluated according to the size and concentration of the PSMPs in experiments where the frequency was varied. Lysis occurred mainly by mechanical collision using PSMPs in a high-frequency domain, and the lysis efficiency was improved by increasing the application time and the energy density of the SAWs. Raman signals from the lysed cells were greatly enhanced by nanogaps formed by the AuNPs, which were evenly distributed irrespective of the SAWs through the frequency-independent behavior of the AuNPs. Finally, detailed Raman spectra of MDA-MB-231, malignant breast cancer cells, were acquired, and various organic matter-derived peaks were observed. The 95% confidence region for cells subjected to lysis was more widely distributed than that of cells not subjected to lysis. The proposed SAW platform is expected to facilitate the detection of small quantities and to be applied in biomedical applications.


Assuntos
Micropartículas Derivadas de Células , Nanopartículas Metálicas , Neoplasias , Ouro , Morte Celular , Poliestirenos
3.
J Chromatogr A ; 1637: 461799, 2021 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-33385744

RESUMO

Dielectrophoresis-field flow fractionation (DEP-FFF) has emerged as an efficient in-vitro, non-invasive, and label-free mechanism to manipulate a variety of nano- and micro-scaled particles in a continuous-flow manner. The technique is mainly used to fractionate particles/cells based on differences in their sizes and/or dielectric properties by employing dielectrophoretic force as an external force field applied perpendicular to the flow direction. The dielectrophoretic force is the result of a spatially non-uniform electric field in the microchannel that can be generated either by exploiting microchannel geometry or using special arrangements of microelectrode arrays. Several two-dimensional (e.g., coplanar interdigitated, castellated) and three-dimensional (e.g., top-bottom, side-wall) microelectrode designs have been successfully utilized to perform fractionation of heterogeneous samples. Although originally introduced as a separation technique, DEP-FFF has attracted increasing interest in performing other important operations such as switching, focusing, dipping, and surface functionalization of target particles. Nonetheless, the technique still suffers from limitations such as low throughput and joule heating. By comparatively analyzing recent developments that address these shortcomings, this work is a step forward towards realizing the full potential of DEP-FFF as an ideal candidate for point-of-care (POC) devices with diverse applications in the fields of biomedical, chemical, and environmental engineering.


Assuntos
Eletroforese/métodos , Fracionamento por Campo e Fluxo/métodos
4.
Med Eng Phys ; 81: 130-135, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32507676

RESUMO

This article details simulation based study of cell separation in a dielectrophoretic microfluidic device. The device consists of a narrow microchannel connected to a wide microchannel with several finite sized planar interdigitated transducer electrodes protruding into the narrow microchannel from one of its sidewalls. In the narrow microchannel, the circulating tumor cells are subjected to positive dielectrophoresis while the regular cells are subjected to negative dielectrophoresis to achieve separation and as all cells move in to the wide microchannel, the physical distance between the two types of cells increases thereby making their collection from the device easier. Equations describing motion, fluid field, electric field, and electric potential form the mathematical model and accounts for forces related to inertia, drag, and dielectrophoresis. Applied electric potential, electrode/gap length, and tumor cell diameter have a positive effect on the performance metrics while velocity of the medium and microchannel width have negative effect on the performance metrics. The model presented in this article is beneficial in realizing liquid biopsy with the desired performance metrics using the proposed microfluidic device.


Assuntos
Separação Celular , Simulação por Computador , Eletroforese , Dispositivos Lab-On-A-Chip , Biópsia Líquida , Técnicas Analíticas Microfluídicas , Células Neoplásicas Circulantes
5.
Analyst ; 144(12): 3853-3860, 2019 Jun 21.
Artigo em Inglês | MEDLINE | ID: mdl-31099354

RESUMO

This article documents the effect of dielectrophoresis on living cells. Given the longer duration procedures performed on microfluidic platforms, the influence of electric fields of high intensity may be of interest in manipulations other than dielectrophoresis. The crossover frequencies of several cell lines were experimentally determined using a microfluidic device. The crossover frequencies are investigated at different medium conductivities for red blood cells, white blood cells-Jurkat, 92.1 and OCM melanoma, and MDA-MB-231 breast cancer cell lines. The effect of dielectrophoresis on the cells at the gene level was also investigated by studying the alteration in gene expressions using microarray analysis. The alterations in genes due to the manipulation of cells at 10 kHz and 100 kHz with a sinusoidal 10 V peak signal for 60 minutes are explored. The two frequencies correspond to negative and positive dielectrophoresis, respectively. The cell line MDA-MB-231 is used as a model for studying the genes in this work. The dielectrophoresis was found to alter genes related to apoptosis, rRNA transcription, cellular respiration, energy production, cellular transcriptional activity, and other cellular functions.


Assuntos
Eletricidade , Eletroforese , Expressão Gênica/fisiologia , Linhagem Celular Tumoral , Condutividade Elétrica , Humanos , Dispositivos Lab-On-A-Chip
6.
J Chromatogr B Analyt Technol Biomed Life Sci ; 1087-1088: 133-137, 2018 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-29734073

RESUMO

This short communication introduces a continuous-flow, dielectrophoresis-based lateral fluid flow fractionation microdevice for detection/isolation of circulating tumor cells in the presence of other haematological cells. The device utilizes two sets of planar interdigitated transducer electrodes micropatterned on top of a glass wafer using standard microfabrication techniques. A microchannel with a single inlet and two outlets, realized in polydimethylsiloxane, is bonded on the glass substrate. The two sets of electrodes slightly protrude into the microchannel. Both of the electrode sets are energized with signals at different frequencies and different operating voltages ensuring that the cancer cells experience positive dielectrophoretic force from one set of the electrodes and negative dielectrophoretic force from the other array. Normal cells experience unequal negative dielectrophoretic forces from opposing sets of electrodes. The resultant dielectrophoretic forces on cancer and normal cells push them to flow towards their designed outlets. Successful isolation of green fluorescent protein-labelled MDA-MB-231 breast cancer cells from regular blood cells, both suspended in a sucrose/dextrose medium, is reported in this work.


Assuntos
Separação Celular/métodos , Eletroforese/métodos , Técnicas Analíticas Microfluídicas/métodos , Células Neoplásicas Circulantes , Linhagem Celular Tumoral , Separação Celular/instrumentação , Eletroforese/instrumentação , Desenho de Equipamento , Humanos , Técnicas Analíticas Microfluídicas/instrumentação
7.
Talanta ; 164: 608-611, 2017 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-28107980

RESUMO

This short communication presents the proof-of-concept of a novel dielectrophoretic lab-on-chip for identifying/separating circulating tumor cells for purposes of liquid biopsy. The device consists of a polydimethylsiloxane layer, containing a microchannel, bonded on a glass substrate that holds two sets of planar interdigitated transducer electrodes. The lab-on-chip is operated at a frequency that enables dielectrophoretic force to sort cells, based on type, along the lateral direction. The operating frequency ensures attraction force toward the electrodes on cancer cells and repulsion force toward the center of the microchannel on other cells. Initial tests for demonstrating proof-of-concept have successfully identified/separated green fluorescent protein-labelled MDA-MB-231 breast cancer cells from a mixture of the same and regular blood cells suspended in low conductivity sucrose/dextrose medium.


Assuntos
Eletroforese/instrumentação , Dispositivos Lab-On-A-Chip , Biópsia Líquida/instrumentação , Linhagem Celular Tumoral , Impedância Elétrica , Humanos
8.
J Sep Sci ; 40(5): 1193-1200, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28035792

RESUMO

We describe the design, microfabrication, and testing of a microfluidic device for the separation of cancer cells based on dielectrophoresis. Cancer cells, specifically green fluorescent protein-labeled MDA-MB-231, are successfully separated from a heterogeneous mixture of the same and normal blood cells. MDA-MB-231 cancer cells are separated with an accuracy that enables precise detection and counting of circulating tumor cells present among normal blood cells. The separation is performed using a set of planar interdigitated transducer electrodes that are deposited on the surface of a glass wafer and slightly protrude into the separation microchannel at one side. The device includes two parts, namely, a glass wafer and polydimethylsiloxane element. The device is fabricated using standard microfabrication techniques. All experiments are conducted with low conductivity sucrose-dextrose isotonic medium. The variation in response between MDA-MB-231 cancer cells and normal cells to a certain band of alternating-current frequencies is used for continuous separation of cells. The fabrication of the microfluidic device, preparation of cells and medium, and flow conditions are detailed. The proposed microdevice can be used to detect and separate malignant cells from heterogeneous mixture of cells for the purpose of early screening for cancer.


Assuntos
Separação Celular , Dispositivos Lab-On-A-Chip , Células Neoplásicas Circulantes , Eletroforese , Desenho de Equipamento , Humanos , Técnicas Analíticas Microfluídicas
9.
Electrophoresis ; 32(11): 1327-36, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21500214

RESUMO

In this paper, a method for continuous flow separation of circulating malignant cells from blood in a microfluidic device using dielectrophoresis is discussed. Separation of MDA231 breast cancer cells after mixing with normal blood cells was achieved with a level of accuracy that enabled precise counting of the malignant cells, separation and eventually, sub-culturing. MDA231 cells were separated from the blood to a daughter channel using two pairs of interdigitated activated comb-like electrode structures. All experiments are performed with conductivity adjusted medium samples. The electrode pairs were positioned divergent and convergent with respect to the flow. The AC signals used in the separation are 20 V peak-to-peak with frequencies of 10-50 kHz. The separation is based on balance of magnitude of the dielectrophoretic force and hydrodynamic force. The difference in response between circulating malignant cells and normal cells at a certain band of alternating current frequencies was used for rapid separation of cancer cells from blood. The significance of these experimental results is discussed in this paper, with detailed reporting on the suspension medium, preparation of cells, flow condition and the fabrication process of the microfluidic chip. The present technique could potentially be applied to identify incident cancer at a stage and size that is not yet detectable by standard diagnostic techniques (imaging and biochemical testing). Alternatively, it may also be used to detect cancer recurrences.


Assuntos
Separação Celular/instrumentação , Eletroforese/instrumentação , Técnicas Analíticas Microfluídicas/instrumentação , Células Neoplásicas Circulantes/patologia , Células Sanguíneas/patologia , Linhagem Celular Tumoral , Separação Celular/métodos , Eletrodos , Humanos , Sistemas Microeletromecânicos/instrumentação , Modelos Biológicos
10.
Nanomedicine ; 6(4): 563-9, 2010 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-20060075

RESUMO

To identify genes affected by single-walled carbon nanotubes (SWCNTs) in human normal lung cells, we compared the gene expression profiles of untreated human normal bronchial epithelial (HNBE) cells to profiles of HNBE cells treated with SWCNTs. A complementary DNA microarray analysis consisting of 54,675 human genes revealed marked changes in the expression of 14,294 genes, with 7,029 genes being upregulated and 7,265 being downregulated. This comprehensive list of genes included those associated with cell cycle, apoptosis, cell survival, cell adhesion and motility, signal transduction, and transcription regulation. Additional analysis of 19 genes using reverse transcription-polymerase chain reaction confirmed the microarray analysis. More specifically, our study demonstrates to our knowledge for the first time, evidence that 9 of the 19 genes (most of which encode cell apoptotic, signal transduction, and transcription regulator products) are upregulated in the SWCNTs-treated HNBE cells as compared with untreated cells, whereas the remaining 10 of the 19 (involved in cell adhesion and motility, cell proliferation, and cell survival) are downregulated in SWCNTs-treated HNBE cells in comparison with untreated controls. These findings provide a large body of information regarding gene expression profiles associated with SWCNTs exposure in human lung bronchial epithelial cells, and also represent a source to investigate the mechanism of the effect of SWCNTs in human normal lung cells. From the clinical editor: In this study, the gene expression profile of human normal bronchial epithelial cells was compared with single-wall carbon nanotubes-treated cells. A cDNA microarray analysis consisting of 54,675 human genes revealed significant changes in the expression of 14,294 genes, with 7,029 genes being up-regulated and 7,265 being down-regulated. This serves as a first step in clarification of mechanisms of action and to investigate toxicity in this model.


Assuntos
Brônquios/metabolismo , Células Epiteliais/metabolismo , Perfilação da Expressão Gênica , Nanotubos de Carbono , Brônquios/efeitos dos fármacos , Proliferação de Células , Regulação para Baixo , Células Epiteliais/citologia , Células Epiteliais/efeitos dos fármacos , Humanos , Nanotubos de Carbono/química , Análise de Sequência com Séries de Oligonucleotídeos
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