Cancer Cell Analyses at the Single Cell-Level Using Electroactive Microwell Array Device

Kobayashi, Makito; Kim, Soo Hyeon; Nakamura, Hiroko; Kaneda, Shohei; Fujii, Teruo

doi:10.1371/journal.pone.0139980

Cited by 25 publications

(32 citation statements)

References 23 publications

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“…Plotting single‐cell data of cell diameter and penetration length for MCF‐10A, MCF‐7, HeLa, and PC3 cells revealed strong linear correlations between cell diameter D cell and penetration length L under different gauge pressures applied to the elasticity microcytometer (Figure a). Ensemble averaged data of cell diameter and penetration length were further analyzed, suggesting that MCF‐10A, MCF‐7, HeLa, and PC3 cells had average diameters of 14.88 ± 0.56 μm, 19.81 ± 0.76 μm, 16.01 ± 0.62 μm, and 19.85 ± 0.69 μm, respectively (Figure b), agreeing well with previous reports . Importantly, MCF‐10A cells could remain trapped inside confining channels of the elasticity microcytometer with the gauge pressure up to 400 Pa, whereas MCF‐7, HeLa, and PC3 cells flew through confining channels with gauge pressure > 200 Pa (Figure c).…”

Section: Resultssupporting

confidence: 93%

Multiparametric Biomechanical and Biochemical Phenotypic Profiling of Single Cancer Cells Using an Elasticity Microcytometer

Chen

et al. 2016

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Deep phenotyping of cancer cells at the single-cell level is of critical importance in the era of precision medicine to advance understanding of the precise relationship between gene mutation and cell phenotype and to elucidate biological nature of tumor heterogeneity and their potential biological and clinical implications. Existing microfluidic single-cell phenotyping tools, albeit their high-throughput, high-resolution operation, are limited to phenotypic measurements of 1 – 2 selected morphological and physiological features of single cells. To address the critical need for multiplexed, informative phenotyping of live single cancer cells, herein we reported a microfluidic elasticity microcytometer for multiparametric biomechanical and biochemical phenotypic profiling of free-floating, live single cancer cells to obtain quantitative information of cell size, cell deformability / stiffness, and expression levels of surface receptors simultaneously for the same single live cancer cells. The elasticity microcytometer was implemented for single-cell measurements and comparisons of four human cell lines with distinct metastatic potentials and derived from different human tissues. An analytical model was developed from first principles for the first time to effectively convert cell deformation and adhesion information of single cancer cells encapsulated inside the elasticity microcytometer to cell deformability / stiffness and surface protein expression. Together, the elasticity microcytometer holds a great promise for comprehensive molecular, cellular, and biomechanical phenotypic profiling of live cancer cells at the single cell level, critical for studying intra-tumor cellular and molecular heterogeneity using low-abundance, clinically relevant human cancer cells.

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Section: Resultssupporting

confidence: 93%

Multiparametric Biomechanical and Biochemical Phenotypic Profiling of Single Cancer Cells Using an Elasticity Microcytometer

Chen

et al. 2016

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“…Kobayashi et al. used three different kinds of biochemical assays (immunostaining, viability/apoptosis, and FISH) for cancer cell discrimination after capturing cells by electro‐activated microwells . Most of published articles relied on injection of expensive biochemical reagents, microscopy, and molecular analysis methods to find cell types and their properties.…”

Section: Introductionmentioning

confidence: 99%

Dielectrophoresis assisted loading and unloading of microwells for impedance spectroscopy

et al. 2017

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Dielectric spectroscopy (DS) is a non-invasive, label-free, fast, and promising technique for measuring dielectric properties of biological cells in real time. We demonstrate a microchip that consists of electro-activated micro-well arrays for positive dielectrophoresis (pDEP) assisted cell capture, DS measurements, and negative dielectrophoresis (nDEP) driven cell unloading; thus, providing a high throughput cell analysis platform. To the best of our knowledge, this is the first microfluidic chip that combines electro-activated micro-wells and DS to analyze biological cells. Device performance is tested using Saccharomyces Cerevisiae (yeast) cells. DEP response of yeast cells is determined by measuring their Clausius-Mossotti (CM) factor using biophysical models in parallel plate micro-electrode geometry. This information is used to determine the excitation frequency to load and unload wells. Effect of yeast cells on the measured impedance spectrum was examined both experimentally and numerically. Good match between the numerical and experimental results establishes the potential use of the micro-chip device for extracting sub-cellular properties of biological cells in a rapid and non-expensive manner.

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“…The occasional trapping of more than one cell in the same well is dependent on the level of the dielectrophoresis trapping force applied in combination with the flow rate and is inevitably happening using a force strong enough to ensure a high trapping efficiency. It was observed that this phenomenon was more frequent in the beginning of the trapping area36. Despite of this it was never challenging to identify and count the trapped cells as the shallow wells will not allow cells to be trapped directly above each other.…”

Section: Resultsmentioning

confidence: 99%

Label-free single-cell separation and imaging of cancer cells using an integrated microfluidic system

Antfolk

Kim

Koizumi

et al. 2017

Sci Rep

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The incidence of cancer is increasing worldwide and metastatic disease, through the spread of circulating tumor cells (CTCs), is responsible for the majority of the cancer deaths. Accurate monitoring of CTC levels in blood provides clinical information supporting therapeutic decision making, and improved methods for CTC enumeration are asked for. Microfluidics has been extensively used for this purpose but most methods require several post-separation processing steps including concentration of the sample before analysis. This induces a high risk of sample loss of the collected rare cells. Here, an integrated system is presented that efficiently eliminates this risk by integrating label-free separation with single cell arraying of the target cell population, enabling direct on-chip tumor cell identification and enumeration. Prostate cancer cells (DU145) spiked into a sample with whole blood concentration of the peripheral blood mononuclear cell (PBMC) fraction were efficiently separated and trapped at a recovery of 76.2 ± 5.9% of the cancer cells and a minute contamination of 0.12 ± 0.04% PBMCs while simultaneously enabling a 20x volumetric concentration. This constitutes a first step towards a fully integrated system for rapid label-free separation and on-chip phenotypic characterization of circulating tumor cells from peripheral venous blood in clinical practice.

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Cancer Cell Analyses at the Single Cell-Level Using Electroactive Microwell Array Device

Cited by 25 publications

References 23 publications

Multiparametric Biomechanical and Biochemical Phenotypic Profiling of Single Cancer Cells Using an Elasticity Microcytometer

Multiparametric Biomechanical and Biochemical Phenotypic Profiling of Single Cancer Cells Using an Elasticity Microcytometer

Dielectrophoresis assisted loading and unloading of microwells for impedance spectroscopy

Label-free single-cell separation and imaging of cancer cells using an integrated microfluidic system

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