Monitoring oxygen consumption of single mouse embryos using an integrated electrochemical microdevice

Date, Yasumoto; Takano, Shoichi; Shiku, Hitoshi; Ino, Kosuke; Ito-Sasaki, Takahiro; Yokoo, Masaki; Abé, Hiroyuki; Matsue, Tomokazu

doi:10.1016/j.bios.2011.08.037

Cited by 43 publications

(34 citation statements)

References 18 publications

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“…The respiratory activity of microtissues is determined by measuring variations in the dissolved oxygen concentration in the solution in the vicinity of the microtissues, and for living microtissues, a gradient in oxygen concentration is expected from the microtissue surface to the bulk of the solution [9], [21]. Therefore, in a first step, the relationship between the oxygen reduction current which is directly proportional to the dissolved oxygen concentration [32], [33], and the distance from the microtissues is determined by scanning over living HeLa microtissues at different heights, along with approach curves over a Petri dish surface and microtissue surface, as shown in Fig. 5B.…”

Section: Resultsmentioning

confidence: 99%

Microstamped Petri Dishes for Scanning Electrochemical Microscopy Analysis of Arrays of Microtissues

et al. 2014

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While scanning electrochemical microscopy (SECM) is a powerful technique for non-invasive analysis of cells, SECM-based assays remain scarce and have been mainly limited so far to single cells, which is mostly due to the absence of suitable platform for experimentation on 3D cellular aggregates or microtissues. Here, we report stamping of a Petri dish with a microwell array for large-scale production of microtissues followed by their in situ analysis using SECM. The platform is realized by hot embossing arrays of microwells (200 μm depth; 400 μm diameter) in commercially available Petri dishes, using a PDMS stamp. Microtissues form spontaneously in the microwells, which is demonstrated here using various cell lines (e.g., HeLa, C2C12, HepG2 and MCF-7). Next, the respiratory activity of live HeLa microtissues is assessed by monitoring the oxygen reduction current in constant height mode and at various distances above the platform surface. Typically, at a 40 μm distance from the microtissue, a 30% decrease in the oxygen reduction current is measured, while above 250 μm, no influence of the presence of the microtissues is detected. After exposure to a model drug (50% ethanol), no such changes in oxygen concentration are found at any height in solution, which reflects that microtissues are not viable anymore. This is furthermore confirmed using conventional live/dead fluorescent stains. This live/dead assay demonstrates the capability of the proposed approach combining SECM and microtissue arrays formed in a stamped Petri dish for conducting cellular assays in a non-invasive way on 3D cellular models.

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Section: Resultsmentioning

confidence: 99%

Microstamped Petri Dishes for Scanning Electrochemical Microscopy Analysis of Arrays of Microtissues

et al. 2014

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“…[43][44][45] In one such device, a single mouse embryo was positioned in a microwell surrounded by electrodes (Fig. 8), and the O 2 consumption of the two-cell, morula, and blastocyst stages was successfully measured.…”

Section: Electrode Array Devicesmentioning

confidence: 99%

“…8 Chip device with three platinum working electrodes, a circular SiO 2 insulator layer, and a cylindrical micropit for evaluation of respiratory activity. 45 Scale bars: 100 µm. Reproduced with permission from Elsevier ©2011.…”

Section: Electrode Array Devicesmentioning

confidence: 99%

Micro/nanoelectrochemical probe and chip devices for evaluation of three-dimensional cultured cells

Ino

Şen

Shiku

et al. 2017

Analyst

Self Cite

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Herein, we present an overview of recent research progress in the development of micro/nanoelectrochemical probe and chip devices for the evaluation of three-dimensional (3D) cultured cells. First, we discuss probe devices: a general outline, evaluation of O 2 consumption, enzyme-modified electrodes, evaluation of endogenous enzyme activity, and the collection of cell components from cell aggregates are discussed. The next section is focused on integrated chip devices: a general outline, electrode array devices, smart electrode array devices, droplet detection of 3D cultured cells, cell manipulation using dielectrophoresis (DEP), and electrodeposited hydrogels used for fabrication of 3D cultured cells on chip devices are discussed. Finally, we provide a summary and discussion of future directions of research in this field.

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“…Microfluidic technology has been utilized to facilitate gamete evaluation (4, 5) to remove cumulus cells and zona pellucida (6–8), for embryo culture (9), for transportation of embryos (10, 11), for monitoring of embryos (12, 13) and for studies on sperm selection and fertilization (14–16). Commonly used methods involve static culture methods (3), microwell designs in which the oocytes are confined (17, 18), and individual chamber based approaches (19).…”

Section: Introductionmentioning

confidence: 99%

Simple perfusion apparatus for manipulation, tracking, and study of oocytes and embryos

Angione

Oulhen

Brayboy

et al. 2015

Fertility and Sterility

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Objective To develop and implement a device and protocol for oocyte analysis at a single cell level. The device must be capable of high resolution imaging, temperature control, perfusion of media, drugs, sperm, and immunolabeling reagents all at defined flow-rates. Each oocyte and resultant embryo must remain spatially separated and defined. Design Experimental laboratory study Setting University and Academic Center for reproductive medicine. Patients/Animals Women with eggs retrieved for ICSI cycles, adult female FVBN and B6C3F1 mouse strains, sea stars. Intervention Real-time, longitudinal imaging of oocytes following fluorescent labeling, insemination, and viability tests. Main outcome measure(s) Cell and embryo viability, immunolabeling efficiency, live cell endocytosis quantitation, precise metrics of fertilization and embryonic development. Results Single oocytes were longitudinally imaged following significant changes in media, markers, endocytosis quantitation, and development, all with supreme control by microfluidics. Cells remained viable, enclosed, and separate for precision measurements, repeatability, and imaging. Conclusions We engineered a simple device to load, visualize, experiment, and effectively record individual oocytes and embryos, without loss of cells. Prolonged incubation capabilities provide longitudinal studies without need for transfer and potential loss of cells. This simple perfusion apparatus (SPA) provides for careful, precise, and flexible handling of precious samples facilitating clinical in vitro fertilization approaches.

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Monitoring oxygen consumption of single mouse embryos using an integrated electrochemical microdevice

Cited by 43 publications

References 18 publications

Microstamped Petri Dishes for Scanning Electrochemical Microscopy Analysis of Arrays of Microtissues

Microstamped Petri Dishes for Scanning Electrochemical Microscopy Analysis of Arrays of Microtissues

Micro/nanoelectrochemical probe and chip devices for evaluation of three-dimensional cultured cells

Simple perfusion apparatus for manipulation, tracking, and study of oocytes and embryos

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