Editorial

3 pages, 163 KiB

Open AccessEditorial

Editorial for the Special Issue on Micro and Nano Devices for Cell Analysis

by Shohei Yamamura

Micromachines 2021, 12(7), 840; https://doi.org/10.3390/mi12070840 - 19 Jul 2021

Cited by 2 | Viewed by 1331

In recent years, miniaturized systems (micro- and nano-devices) called a lab-on-a-chip or micro-total analysis system (µ-TAS) have received attention as new systems for chemical and biochemical analyses [...] Full article

(This article belongs to the Special Issue Micro and Nano Devices for Cell Analysis)

Research

Jump to: Editorial, Review

10 pages, 3298 KiB

Open AccessArticle

Template Stripping Method-Based Au Nanoarray for Surface-Enhanced Raman Scattering Detection of Antiepileptic Drug

by Tatsuro Endo, Hirotaka Yamada and Kenji Yamada

Micromachines 2020, 11(10), 936; https://doi.org/10.3390/mi11100936 - 14 Oct 2020

Cited by 7 | Viewed by 2501

Abstract

Surface-enhanced Raman scattering (SERS) is a potential candidate for highly sensitive detection of target molecules. A SERS active substrate with a noble metal nanostructure is required for this. However, a SERS active substrate requires complicated fabrication procedures. This in turn makes it difficult [...] Read more.

Surface-enhanced Raman scattering (SERS) is a potential candidate for highly sensitive detection of target molecules. A SERS active substrate with a noble metal nanostructure is required for this. However, a SERS active substrate requires complicated fabrication procedures. This in turn makes it difficult to fabricate highly sensitive SERS active substrates with high reproducibility. To overcome this difficulty, a plasmonic crystal (PC) with periodic noble metal nanostructures was fabricated via the template-stripping method using a polymer-based template. Using SERS active substrates, SERS was successfully achieved using the PC by detecting low concentrations of phenobarbital which is an antiepileptic drug using a commercially available portable Raman module. The PC can be fabricated by demolding the deposited gold layer from a polymer-based template. This method is rapid, economic, and has high reproducibility. SERS can be achieved easily using this PC for a wide variety of applications such as medical, pharmaceutical, and environmental protection. Full article

(This article belongs to the Special Issue Micro and Nano Devices for Cell Analysis)

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10 pages, 18929 KiB

Open AccessFeature PaperEditor’s ChoiceArticle

Microfluidic Separation of Blood Cells Based on the Negative Dielectrophoresis Operated by Three Dimensional Microband Electrodes

by Tomoyuki Yasukawa, Junko Yamada, Hitoshi Shiku, Tomokazu Matsue and Masato Suzuki

Micromachines 2020, 11(9), 833; https://doi.org/10.3390/mi11090833 - 31 Aug 2020

Cited by 8 | Viewed by 2976

Abstract

A microfluidic device is presented for the continuous separation of red blood cells (RBCs) and white blood cells (WBCs) in a label-free manner based on negative dielectrophoresis (n-DEP). An alteration of the electric field, generated by pairs of slanted electrodes (separators) that is [...] Read more.

A microfluidic device is presented for the continuous separation of red blood cells (RBCs) and white blood cells (WBCs) in a label-free manner based on negative dielectrophoresis (n-DEP). An alteration of the electric field, generated by pairs of slanted electrodes (separators) that is fabricated by covering parts of single slanted electrodes with an insulating layer is used to separate cells by their sizes. The repulsive force of n-DEP formed by slanted electrodes prepared on both the top and bottom substrates led to the deflection of the cell flow in lateral directions. The presence of gaps covered with an insulating layer for the electric field on the electrodes allows the passing of RBCs through gaps, while relatively large WBCs (cultured cultured human acute monocytic leukemia cell line (THP-1 cells)) flowed along the slanted separator without passing through the gaps and arrived at an edge in the channel. The passage efficiency for RBCs through the gaps and the arrival efficiency for THP-1 cells to the upper edge in the channel were estimated and found to be 91% and 93%, respectively. Full article

(This article belongs to the Special Issue Micro and Nano Devices for Cell Analysis)

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11 pages, 2631 KiB

Open AccessArticle

Photo-Cleavable Peptide-Poly(Ethylene Glycol) Conjugate Surfaces for Light-Guided Control of Cell Adhesion

by Satoshi Yamaguchi, Yumi Takasaki, Shinya Yamahira and Teruyuki Nagamune

Micromachines 2020, 11(8), 762; https://doi.org/10.3390/mi11080762 - 08 Aug 2020

Cited by 8 | Viewed by 2846

Abstract

Photo-responsive cell attachment surfaces can simplify patterning and recovery of cells in microdevices for medicinal and pharmaceutical research. We developed a photo-responsive surface for controlling the attachment and release of adherent cells on a substrate under light-guidance. The surface comprises a poly(ethylene glycol) [...] Read more.

Photo-responsive cell attachment surfaces can simplify patterning and recovery of cells in microdevices for medicinal and pharmaceutical research. We developed a photo-responsive surface for controlling the attachment and release of adherent cells on a substrate under light-guidance. The surface comprises a poly(ethylene glycol) (PEG)-based photocleavable material that can conjugate with cell-adhesive peptides. Surface-bound peptides were released by photocleavage in the light-exposed region, where the cell attachment was subsequently suppressed by the exposed PEG. Simultaneously, cells selectively adhered to the peptide surface at the unexposed microscale region. After culture, the adhered and spread cells were released by exposure to a light with nontoxic dose level. Thus, the present surface can easily create both cell-adhesive and non-cell-adhesive regions on the substrate by single irradiation of the light pattern, and the adhered cells were selectively released from the light-exposed region on the cell micropattern without damage. This study shows that the photo-responsive surface can serve as a facile platform for the remote-control of patterning and recovery of adherent cells in microdevices. Full article

(This article belongs to the Special Issue Micro and Nano Devices for Cell Analysis)

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14 pages, 5613 KiB

Open AccessFeature PaperEditor’s ChoiceArticle

Facile Fabrication of Thin-Bottom Round-Well Plates Using the Deformation of PDMS Molds and Their Application for Single-Cell PCR

by Shinya Yamahira and Yuji Heike

Micromachines 2020, 11(8), 748; https://doi.org/10.3390/mi11080748 - 31 Jul 2020

Cited by 2 | Viewed by 5402

Abstract

Recently, microdevices made of resins have been strongly supporting cell analysis in a range of fields, from fundamental life science research to medical applications. Many microdevices are fabricated by molding resin to a mold made precisely from rigid materials. However, because dimensional errors [...] Read more.

Recently, microdevices made of resins have been strongly supporting cell analysis in a range of fields, from fundamental life science research to medical applications. Many microdevices are fabricated by molding resin to a mold made precisely from rigid materials. However, because dimensional errors in the mold are also accurately printed to the products, the accuracy of the product is limited to less than the accuracy of the rigid mold. Therefore, we hypothesized that if dimensional errors could be self-corrected by elastic molds, microdevices could be facilely fabricated with precision beyond that of molds. In this paper, we report a novel processing strategy in which an elastic mold made of polymethylsiloxane (PDMS) deforms to compensate for the dimensional error on the products. By heat-press molding a polycarbonate plate using a mold that has 384 PDMS convexes with a large dimensional error of height of ± 15.6 µm in standard deviation, a 384-round-well plate with a bottom thickness 13.3 ± 2.3 µm (n = 384) was easily fabricated. Finally, single-cell observation and polymerase chain reactions (PCRs) demonstrated the application of the products made by elastic PDMS molds. Therefore, this processing method is a promising strategy for facile, low-cost, and higher precision microfabrication. Full article

(This article belongs to the Special Issue Micro and Nano Devices for Cell Analysis)

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12 pages, 1071 KiB

Open AccessFeature PaperArticle

Analysis of Single Nucleotide-Mutated Single-Cancer Cells Using the Combined Technologies of Single-Cell Microarray Chips and Peptide Nucleic Acid-DNA Probes

by Hajime Shigeto, Eriko Yamada, Mizuki Kitamatsu, Takashi Ohtsuki, Akira Iizuka, Yasuto Akiyama and Shohei Yamamura

Micromachines 2020, 11(7), 628; https://doi.org/10.3390/mi11070628 - 27 Jun 2020

Cited by 11 | Viewed by 2699

Abstract

Research into cancer cells that harbor gene mutations relating to anticancer drug-resistance at the single-cell level has focused on the diagnosis of, or treatment for, cancer. Several methods have been reported for detecting gene-mutated cells within a large number of non-mutated cells; however, [...] Read more.

Research into cancer cells that harbor gene mutations relating to anticancer drug-resistance at the single-cell level has focused on the diagnosis of, or treatment for, cancer. Several methods have been reported for detecting gene-mutated cells within a large number of non-mutated cells; however, target single nucleotide-mutated cells within a large number of cell samples, such as cancer tissue, are still difficult to analyze. In this study, a new system is developed to detect and isolate single-cancer cells expressing the T790M-mutated epidermal growth factor receptor (EGFR) mRNA from multiple non-mutated cancer cells by combining single-cell microarray chips and peptide nucleic acid (PNA)-DNA probes. The single-cell microarray chip is made of polystyrene with 62,410 microchambers (31-40 µm diameter). The T790M-mutated lung cancer cell line, NCI-H1975, and non-mutated lung cancer cell line, A549, were successfully separated into single cells in each microchambers on the chip. Only NCI-H1975 cell was stained on the chip with a fluorescein isothiocyanate (FITC)-conjugated PNA probe for specifically detecting T790M mutation. Of the NCI-H1975 cells that spiked into A549 cells, 0–20% were quantitatively analyzed within 1 h, depending on the spike concentration. Therefore, our system could be useful in analyzing cancer tissue that contains a few anticancer drug-resistant cells. Full article

(This article belongs to the Special Issue Micro and Nano Devices for Cell Analysis)

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13 pages, 3796 KiB

Open AccessArticle

Multi-Color Enhanced Fluorescence Imaging of a Breast Cancer Cell with A Hole-Arrayed Plasmonic Chip

by Makiko Yoshida, Hinako Chida, Fukiko Kimura, Shohei Yamamura and Keiko Tawa

Micromachines 2020, 11(6), 604; https://doi.org/10.3390/mi11060604 - 22 Jun 2020

Cited by 10 | Viewed by 2786

Abstract

Breast cancer cells of MDA-MB-231 express various types of membrane proteins in the cell membrane. In this study, two types of membrane proteins in MDA-MB-231 cells were observed using a plasmonic chip with an epifluorescence microscope. The targeted membrane proteins were epithelial cell [...] Read more.

Breast cancer cells of MDA-MB-231 express various types of membrane proteins in the cell membrane. In this study, two types of membrane proteins in MDA-MB-231 cells were observed using a plasmonic chip with an epifluorescence microscope. The targeted membrane proteins were epithelial cell adhesion molecules (EpCAMs) and epidermal growth factor receptor (EGFR), and Alexa^®488-EGFR antibody and allophycocyanin (APC)-labeled EpCAM antibody were applied to the fluorescent detection. The plasmonic chip used in this study is composed of a two-dimensional hole-array structure, which is expected to enhance the fluorescence at different resonance wavelengths due to two kinds of grating pitches in a square side and a diagonal direction. As a result of multi-color imaging, the enhancement factor of Alexa^®488-EGFR and APC-EpCAM was 13 ± 2 and 12 ± 2 times greater on the plasmonic chip, respectively. The excited wavelength or emission wavelength of each fluorescent agent is due to consistency with plasmon resonance wavelength in the hole-arrayed chip. The multi-color fluorescence images of breast cancer cells were improved by the hole-arrayed plasmonic chip. Full article

(This article belongs to the Special Issue Micro and Nano Devices for Cell Analysis)

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14 pages, 5975 KiB

Open AccessArticle

Cell Culture on Low-Fluorescence and High-Resolution Photoresist

by Hidetaka Ueno, Katsuya Maruo, Masatoshi Inoue, Hidetoshi Kotera and Takaaki Suzuki

Micromachines 2020, 11(6), 571; https://doi.org/10.3390/mi11060571 - 04 Jun 2020

Cited by 7 | Viewed by 2574

Abstract

2D and 3D topographic cues made of photoresist, a polymer, are used for cell culture and cell analysis. Photoresists used for cell analysis provide the surface conditions necessary for proper cell growth, along with patterning properties of a wide range and high precision, [...] Read more.

2D and 3D topographic cues made of photoresist, a polymer, are used for cell culture and cell analysis. Photoresists used for cell analysis provide the surface conditions necessary for proper cell growth, along with patterning properties of a wide range and high precision, and low auto-fluorescence that does not affect fluorescence imaging. In this study, we developed a thick negative photoresist SJI-001 possessing the aforementioned properties. We evaluated the surface conditions of SJI-001 affecting cell culture. First, we studied the wettability of SJI-001, which was changed by plasma treatment, conducted as a pretreatment on a plastic substrate before cell seeding. SJI-001 was more chemically stable than SU-8 used for fabricating the micro-electromechanical systems (MEMS). Furthermore, the doubling time and adhesion rate of adherent HeLa cells cultured on untreated SJI-001 were 25.2 h and 74%, respectively, thus indicating its suitability for cell culture over SU-8. In addition, we fabricated a cell culture plate with a 3D lattice structure, three micrometers in size, using SJI-001. HeLa cells seeded on this plate remained attached over five days. Therefore, SJI-001 exhibits surface conditions suitable for cell culture and has several bioapplications including microstructures and cell chips for cell culture and cell analysis. Full article

(This article belongs to the Special Issue Micro and Nano Devices for Cell Analysis)

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9 pages, 1946 KiB

Open AccessArticle

Simultaneously Quantifying Both Young’s Modulus and Specific Membrane Capacitance of Bladder Cancer Cells with Different Metastatic Potential

by Na Liu, Mengying Leng, Tao Yue, Liang Dong, Yuanyuan Liu, Yan Peng, Huayan Pu, Shaorong Xie and Jun Luo

Micromachines 2020, 11(3), 249; https://doi.org/10.3390/mi11030249 - 27 Feb 2020

Cited by 3 | Viewed by 2302

Abstract

Both Young’s modulus and specific membrane capacitance (SMC) are two important physical parameters for characterizing cell status. In this paper, we utilized a thin-neck-micropipette aspiration system to simultaneously quantify Young’s modulus and SMC value of six types of cell lines in different progression [...] Read more.

Both Young’s modulus and specific membrane capacitance (SMC) are two important physical parameters for characterizing cell status. In this paper, we utilized a thin-neck-micropipette aspiration system to simultaneously quantify Young’s modulus and SMC value of six types of cell lines in different progression grades, which include four grades from the lowest metastatic potential G1 to the highest potential G4. We investigated how these two physical properties possess heterogeneities in bladder cancer cells with different grades and what roles they might play in grading bladder cancer. The characterization results of these cells of different cancer grades is linearly correlated with the cancer grades, showing that the Young’s modulus is negatively linearly correlated with bladder cancer grades, while SMC shows a positive linear correlation. Furthermore, the combination of these two physical properties on a scatter diagram clearly shows the cell groups with different cancer grades, which means that this combination could be a potential tumor grading marker to identify cancer cells with different metastatic potential. Full article

(This article belongs to the Special Issue Micro and Nano Devices for Cell Analysis)

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16 pages, 3865 KiB

Open AccessFeature PaperArticle

Single Cell Analysis of Neutrophils NETs by Microscopic LSPR Imaging System

by Riyaz Ahmad Mohamed Ali, Daiki Mita, Wilfred Espulgar, Masato Saito, Masayuki Nishide, Hyota Takamatsu, Hiroyuki Yoshikawa and Eiichi Tamiya

Micromachines 2020, 11(1), 52; https://doi.org/10.3390/mi11010052 - 31 Dec 2019

Cited by 10 | Viewed by 2986

Abstract

A simple microengraving cell monitoring method for neutrophil extracellular traps (NETs) released from single neutrophils has been realized using a polydimethylsiloxane (PDMS) microwell array (MWA) sheet on a plasmon chip platform. An imbalance between NETs formation and the succeeding degradation (NETosis) are considered [...] Read more.

A simple microengraving cell monitoring method for neutrophil extracellular traps (NETs) released from single neutrophils has been realized using a polydimethylsiloxane (PDMS) microwell array (MWA) sheet on a plasmon chip platform. An imbalance between NETs formation and the succeeding degradation (NETosis) are considered associated with autoimmune disease and its pathogenesis. Thus, an alternative platform that can conduct monitoring of this activity on single cell level at minimum cost but with great sensitivity is greatly desired. The developed MWA plasmon chips allow single cell isolation of neutrophils from 150 µL suspension (6.0 × 10⁵ cells/mL) with an efficiency of 36.3%; 105 microwells with single cell condition. To demonstrate the utility of the chip, trapped cells were incubated between 2 to 4 h after introducing with 100 nM phorbol 12-myristate 13-acetate (PMA) before measurement. Under observation using a hyperspectral imaging system that allows high-throughput screening, the neutrophils stimulated by PMA solution show a significant release of fibrils and NETs after 4 h, with observed maximum areas between 314–758 µm². An average absorption peak wavelength shows a redshift of Δλ = 1.5 nm as neutrophils release NETs. Full article

(This article belongs to the Special Issue Micro and Nano Devices for Cell Analysis)

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10 pages, 1926 KiB

Open AccessArticle

Mechanical Property Changes in Breast Cancer Cells Induced by Stimulation with Macrophage Secretions in Vitro

by Hyonchol Kim, Kenta Ishibashi, Tomoko Okada and Chikashi Nakamura

Micromachines 2019, 10(11), 738; https://doi.org/10.3390/mi10110738 - 30 Oct 2019

Cited by 9 | Viewed by 2765

Abstract

The contribution of secretions from tumor-associated macrophage (TAM)-like cells to the stimulation of mechanical property changes in murine breast cancer cells was studied using an in vitro model system. A murine breast cancer cell line (FP10SC2) was stimulated by adding macrophage (J774.2) cultivation [...] Read more.

The contribution of secretions from tumor-associated macrophage (TAM)-like cells to the stimulation of mechanical property changes in murine breast cancer cells was studied using an in vitro model system. A murine breast cancer cell line (FP10SC2) was stimulated by adding macrophage (J774.2) cultivation medium containing stimulation molecules secreted from the macrophages, and changes in mechanical properties were compared before and after stimulation. As a result, cell elasticity decreased, degradation ability of the extracellular matrix increased, and the expression of plakoglobin was upregulated. These results indicate that cancer cell malignancy is upregulated by this stimulation. Moreover, changes in intercellular adhesion strengths between pairs of cancer cells were measured before and after stimulation using atomic force microscopy (AFM). The maximum force required to separate cells was increased by stimulation with the secreted factors. These results indicate the possibility that TAMs cause changes in the mechanical properties of cancer cells in tumor microenvironments, and in vitro measurements of mechanical property changes in cancer cells will be useful to study interactions between cells in tumor microenvironments. Full article

(This article belongs to the Special Issue Micro and Nano Devices for Cell Analysis)

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Review

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23 pages, 5807 KiB

Open AccessReview

Recent Advances in Monitoring Cell Behavior Using Cell-Based Impedance Spectroscopy

by Qusai Hassan, Soha Ahmadi and Kagan Kerman

Micromachines 2020, 11(6), 590; https://doi.org/10.3390/mi11060590 - 13 Jun 2020

Cited by 33 | Viewed by 4820

Abstract

Cell-based impedance spectroscopy (CBI) is a powerful tool that uses the principles of electrochemical impedance spectroscopy (EIS) by measuring changes in electrical impedance relative to a voltage applied to a cell layer. CBI provides a promising platform for the detection of several properties [...] Read more.

Cell-based impedance spectroscopy (CBI) is a powerful tool that uses the principles of electrochemical impedance spectroscopy (EIS) by measuring changes in electrical impedance relative to a voltage applied to a cell layer. CBI provides a promising platform for the detection of several properties of cells including the adhesion, motility, proliferation, viability and metabolism of a cell culture. This review gives a brief overview of the theory, instrumentation, and detection principles of CBI. The recent applications of the technique are given in detail for research into cancer, neurodegenerative diseases, toxicology as well as its application to 2D and 3D in vitro cell cultures. CBI has been established as a biophysical marker to provide quantitative cellular information, which can readily be adapted for single-cell analysis to complement the existing biomarkers for clinical research on disease progression. Full article

(This article belongs to the Special Issue Micro and Nano Devices for Cell Analysis)

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23 pages, 4225 KiB

Open AccessFeature PaperReview

Recent Advances in Electrochemiluminescence-Based Systems for Mammalian Cell Analysis

by Kaoru Hiramoto, Elena Villani, Tomoki Iwama, Keika Komatsu, Shinsuke Inagi, Kumi Y. Inoue, Yuji Nashimoto, Kosuke Ino and Hitoshi Shiku

Micromachines 2020, 11(5), 530; https://doi.org/10.3390/mi11050530 - 22 May 2020

Cited by 37 | Viewed by 5026

Abstract

Mammalian cell analysis is essential in the context of both fundamental studies and clinical applications. Among the various techniques available for cell analysis, electrochemiluminescence (ECL) has attracted significant attention due to its integration of both electrochemical and spectroscopic methods. In this review, we [...] Read more.

Mammalian cell analysis is essential in the context of both fundamental studies and clinical applications. Among the various techniques available for cell analysis, electrochemiluminescence (ECL) has attracted significant attention due to its integration of both electrochemical and spectroscopic methods. In this review, we summarize recent advances in the ECL-based systems developed for mammalian cell analysis. The review begins with a summary of the developments in luminophores that opened the door to ECL applications for biological samples. Secondly, ECL-based imaging systems are introduced as an emerging technique to visualize single-cell morphologies and intracellular molecules. In the subsequent section, the ECL sensors developed in the past decade are summarized, the use of which made the highly sensitive detection of cell-derived molecules possible. Although ECL immunoassays are well developed in terms of commercial use, the sensing of biomolecules at a single-cell level remains a challenge. Emphasis is therefore placed on ECL sensors that directly detect cellular molecules from small portions of cells or even single cells. Finally, the development of bipolar electrode devices for ECL cell assays is introduced. To conclude, the direction of research in this field and its application prospects are described. Full article

(This article belongs to the Special Issue Micro and Nano Devices for Cell Analysis)

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