Cell Microarray Technologies for High-Throughput Cell-Based Biosensors

Hong, Hye Jin; Koom, Woong Sub; Koh, Won Gun

doi:10.3390/s17061293

Cited by 40 publications

(17 citation statements)

References 199 publications

(232 reference statements)

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“…In particular, electric cell-substrate impedance sensing (ECIS) has been effectively used to assess bioviability, demonstrating a potential for high-throughput monitoring of cells adhesion and proliferation in vitro. [4][5][6][7] However, these systems are generally not compatible with the microscopy techniques extensively used for life-sciences research, the current gold standard for cell physiology monitoring, because of either the opaque nature of the substrates and/or of the electrodes used for sensing. The excellent works by Róisín Owens and collaborators have in part overcome the limitations of standard ECIS, demonstrating organic electrochemical transistors able to detect and measure cell adhesion, proliferation and detachment in vitro with enhanced sensitivity and temporal resolution compared to standard technologies, with the added advantage of allowing for simultaneous electrical and optical analyses.…”

Section: Introductionmentioning

confidence: 99%

Real-Time Monitoring of Cellular Cultures with Electrolyte-Gated Carbon Nanotube Transistors

Scuratti

Bonacchini

Bossio

et al. 2019

ACS Appl. Mater. Interfaces

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Cell-based biosensors constitute a fundamental tool in biotechnology, and their relevance has greatly increased in recent years as a result of a surging demand for reduced animal testing and for high-throughput and cost-effective in vitro screening platforms dedicated to environmental and biomedical diagnostics, drug development and toxicology. In this context, electrochemical/electronic cell-based biosensors represent a promising class of devices that enable long-term and real-time monitoring of cell physiology in a non-invasive and label-free fashion, with a remarkable potential for process automation and parallelization. Common limitations of this class of devices at large include the need for substrate surface modification strategies to ensure cell adhesion and immobilization, limited compatibility with complementary optical cell-probing techniques, and need for frequency-dependent measurements, which rely on elaborated equivalent electrical circuit models for data analysis and interpretation. We hereby demonstrate the monitoring of cell adhesion and detachment through the time-dependent variations in the quasi-static characteristic current curves of a highly stable electrolyte-gated transistor, based on an optically transparent network of printable polymer-wrapped semiconducting carbon-nanotubes. MAIN TEXTOptical cell viability assay and immunofluorescence staining: The proliferation was evaluated after 1, 2, 3, and 4 d in vitro. For each time point the medium was removed and replaced with RPMI without phenol red containing 0.5 mg mL −1 of MTT reagent (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, Sigma-Aldrich). Cells were reincubated at 37 °C for 3 h. Formazan salt produced by cells through reduction of MTT was then solubilized with 200 mL of ethanol and the absorbance was read at 560 and 690 nm (using a microplate reader TECAN Spark10M). The proliferation cell rate was calculated as the difference in absorbed intensity at 560 and 690 nm. Cells grown on glass coverslips coated with SWCNT networks were washed twice with PBS and fixed for 15 min at RT in 4 % paraformaldehyde and 4 % sucrose in 0.12 M sodium phosphate buffer, pH 7.4. Fixed cells were pre-incubated for 20 min in gelatin dilution buffer (GDB: 0.02 M sodium phosphate buffer, pH 7.4, 0.45 M NaCl, 0.2% (w/v) gelatin) containing 0.3% (v/v)

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Section: Introductionmentioning

confidence: 99%

Real-Time Monitoring of Cellular Cultures with Electrolyte-Gated Carbon Nanotube Transistors

Scuratti

Bonacchini

Bossio

et al. 2019

ACS Appl. Mater. Interfaces

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“…As cell collections grow, further miniaturization of cell assays is needed to increase parallelism of the analyses. Cell microarrays provide an attractive solution, as they could increase the throughput significantly [1][2][3]. A cellular microarray consists of a solid support wherein small volumes of different biomolecules and cells can be displayed in defined locations, allowing the multiplexed interrogation of living cells and the analysis of cellular responses [4,5].…”

Section: Introductionmentioning

confidence: 99%

Robotic Cell Printing for Constructing Living Yeast Cell Microarrays in Microfluidic Chips

2020

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Living cell microarrays in microfluidic chips allow the non-invasive multiplexed molecular analysis of single cells. Here, we developed a simple and affordable perfusion microfluidic chip containing a living yeast cell array composed of a population of cell variants (green fluorescent protein (GFP)-tagged Saccharomyces cerevisiae clones). We combined mechanical patterning in 102 microwells and robotic piezoelectric cell dispensing in the microwells to construct the cell arrays. Robotic yeast cell dispensing of a yeast collection from a multiwell plate to the microfluidic chip microwells was optimized. The developed microfluidic chip and procedure were validated by observing the growth of GFP-tagged yeast clones that are linked to the cell cycle by time-lapse fluorescence microscopy over a few generations. The developed microfluidic technology has the potential to be easily upscaled to a high-density cell array allowing us to perform dynamic proteomics and localizomics experiments.

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“…Micropatterned protein or DNA substrates have been successfully used to address many different biological questions in various research areas, including disease diagnosis, clinical and pharmacogenomics research, the analysis of cellular functions, and drug discovery [1][2][3][4]. In this regard, micropatterned surfaces as part of lab-on-a-chip systems are of special interest as they can fulfill the rising demand for high-throughput diagnostic tools.…”

Section: Introductionmentioning

confidence: 99%

Fabrication, Characterization and Application of Biomolecule Micropatterns on Cyclic Olefin Polymer (COP) Surfaces with Adjustable Contrast

Hager

Haselgrübler²,

Haas³

et al. 2019

Biosensors

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Peptide and protein micropatterns are powerful tools for the investigation of various cellular processes, including protein-protein interactions (PPIs). Within recent years, various approaches for the production of functional surfaces have been developed. Most of these systems use glass as a substrate, which has several drawbacks, including high fragility and costs, especially if implemented for fluorescence microscopy. In addition, conventional fabrication technologies such as microcontact printing (µCP) are frequently used for the transfer of biomolecules to the glass surface. In this case, it is challenging to adjust the biomolecule density. Here, we show that cyclic olefin polymer (COP) foils, with their encouraging properties, including the ease of manufacturing, chemical resistance, biocompatibility, low water absorption, and optical clarity, are a promising alternative to glass substrates for the fabrication of micropatterns. Using a photolithography-based approach, we generated streptavidin/biotinylated antibody patterns on COPs with the possibility of adjusting the pattern contrast by varying plasma activation parameters. Our experimental setup was finally successfully implemented for the analysis of PPIs in the membranes of live cells via total internal reflection fluorescence (TIRF) microscopy.

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Cell Microarray Technologies for High-Throughput Cell-Based Biosensors

Cited by 40 publications

References 199 publications

Real-Time Monitoring of Cellular Cultures with Electrolyte-Gated Carbon Nanotube Transistors

Real-Time Monitoring of Cellular Cultures with Electrolyte-Gated Carbon Nanotube Transistors

Robotic Cell Printing for Constructing Living Yeast Cell Microarrays in Microfluidic Chips

Fabrication, Characterization and Application of Biomolecule Micropatterns on Cyclic Olefin Polymer (COP) Surfaces with Adjustable Contrast

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