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Bipolar Clark-Type Oxygen Electrode Arrays for Imaging and Multiplexed Measurements of the Respiratory Activity of Cells

Various miniature Clark-type oxygen electrodes (COEs), which are typically used to measure dissolved oxygen (DO) concentration in cellular respiration, have been developed since the 1980s. Arrays with individually addressable electrodes that constitute the sensor were used for various applications....

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Main Authors:	Shirato, Yusuke, Hsueh, An-Ju, Ab Mutalib, Nurul Asyikeen, Deng, Yi, Suematsu, Ryohei, Kato, Azusa, Kearney, Bradley M., Kinoshita, Manabu, Suzuki, Hiroaki
Format:	Article
Published:	American Chemical Society 2024
Subjects:	Q Science (General) R Medicine (General)
Online Access:	http://eprints.um.edu.my/45601/ https://doi.org/10.1021/acsomega.3c09802
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spelling	my.um.eprints.456012024-11-06T01:33:35Z http://eprints.um.edu.my/45601/ Bipolar Clark-Type Oxygen Electrode Arrays for Imaging and Multiplexed Measurements of the Respiratory Activity of Cells Shirato, Yusuke Hsueh, An-Ju Ab Mutalib, Nurul Asyikeen Deng, Yi Suematsu, Ryohei Kato, Azusa Kearney, Bradley M. Kinoshita, Manabu Suzuki, Hiroaki Q Science (General) R Medicine (General) Various miniature Clark-type oxygen electrodes (COEs), which are typically used to measure dissolved oxygen (DO) concentration in cellular respiration, have been developed since the 1980s. Arrays with individually addressable electrodes that constitute the sensor were used for various applications. However, the large number of leads and contact pads required for connecting the electrodes and the external instrument complicate the electrode layout and make the operation of integrated COE arrays challenging. Here, we fabricated closed bipolar electrochemical systems comprising 6 x 8 and 4 x 4 arrays of COEs for imaging and multiplexed detection. The cathodic compartment was sealed with a hydrophobic oxygen-permeable membrane to separate the internal electrolyte solution from the sample solutions. Using the bipolar Clark-type oxygen electrode (BCOE) arrays and electrochemiluminescence (ECL), we measured the DO concentration at each cathode. The results revealed that the ECL intensity changed linearly with the DO concentration. In addition, we used ECL imaging to investigate the respiratory activity of Escherichia coli (E. coli) and Pseudomonas aeruginosa (P. aeruginosa) in suspensions with different cell densities. The ECL images showed that the ECL intensity changed noticeably with the bacterial density. The bacterial respiratory activity was then qualitatively analyzed based on the ECL images acquired successively over a time duration. Further, we measured the antibiotic efficacy of piperacillin, oxacillin, gentamicin, and cefmetazole against E. coli and P. aeruginosa using the BCOE. We found that the ECL intensity increased with the antibiotic concentration, thus indicating the suppression of the bacterial respiratory activity. American Chemical Society 2024-02 Article PeerReviewed Shirato, Yusuke and Hsueh, An-Ju and Ab Mutalib, Nurul Asyikeen and Deng, Yi and Suematsu, Ryohei and Kato, Azusa and Kearney, Bradley M. and Kinoshita, Manabu and Suzuki, Hiroaki (2024) Bipolar Clark-Type Oxygen Electrode Arrays for Imaging and Multiplexed Measurements of the Respiratory Activity of Cells. ACS Omega, 9 (9). pp. 10825-10833. ISSN 2470-1343, DOI https://doi.org/10.1021/acsomega.3c09802 <https://doi.org/10.1021/acsomega.3c09802>. https://doi.org/10.1021/acsomega.3c09802 10.1021/acsomega.3c09802
institution	Universiti Malaya
building	UM Library
collection	Institutional Repository
continent	Asia
country	Malaysia
content_provider	Universiti Malaya
content_source	UM Research Repository
url_provider	http://eprints.um.edu.my/
topic	Q Science (General) R Medicine (General)
spellingShingle	Q Science (General) R Medicine (General) Shirato, Yusuke Hsueh, An-Ju Ab Mutalib, Nurul Asyikeen Deng, Yi Suematsu, Ryohei Kato, Azusa Kearney, Bradley M. Kinoshita, Manabu Suzuki, Hiroaki Bipolar Clark-Type Oxygen Electrode Arrays for Imaging and Multiplexed Measurements of the Respiratory Activity of Cells
description	Various miniature Clark-type oxygen electrodes (COEs), which are typically used to measure dissolved oxygen (DO) concentration in cellular respiration, have been developed since the 1980s. Arrays with individually addressable electrodes that constitute the sensor were used for various applications. However, the large number of leads and contact pads required for connecting the electrodes and the external instrument complicate the electrode layout and make the operation of integrated COE arrays challenging. Here, we fabricated closed bipolar electrochemical systems comprising 6 x 8 and 4 x 4 arrays of COEs for imaging and multiplexed detection. The cathodic compartment was sealed with a hydrophobic oxygen-permeable membrane to separate the internal electrolyte solution from the sample solutions. Using the bipolar Clark-type oxygen electrode (BCOE) arrays and electrochemiluminescence (ECL), we measured the DO concentration at each cathode. The results revealed that the ECL intensity changed linearly with the DO concentration. In addition, we used ECL imaging to investigate the respiratory activity of Escherichia coli (E. coli) and Pseudomonas aeruginosa (P. aeruginosa) in suspensions with different cell densities. The ECL images showed that the ECL intensity changed noticeably with the bacterial density. The bacterial respiratory activity was then qualitatively analyzed based on the ECL images acquired successively over a time duration. Further, we measured the antibiotic efficacy of piperacillin, oxacillin, gentamicin, and cefmetazole against E. coli and P. aeruginosa using the BCOE. We found that the ECL intensity increased with the antibiotic concentration, thus indicating the suppression of the bacterial respiratory activity.
format	Article
author	Shirato, Yusuke Hsueh, An-Ju Ab Mutalib, Nurul Asyikeen Deng, Yi Suematsu, Ryohei Kato, Azusa Kearney, Bradley M. Kinoshita, Manabu Suzuki, Hiroaki
author_facet	Shirato, Yusuke Hsueh, An-Ju Ab Mutalib, Nurul Asyikeen Deng, Yi Suematsu, Ryohei Kato, Azusa Kearney, Bradley M. Kinoshita, Manabu Suzuki, Hiroaki
author_sort	Shirato, Yusuke
title	Bipolar Clark-Type Oxygen Electrode Arrays for Imaging and Multiplexed Measurements of the Respiratory Activity of Cells
title_short	Bipolar Clark-Type Oxygen Electrode Arrays for Imaging and Multiplexed Measurements of the Respiratory Activity of Cells
title_full	Bipolar Clark-Type Oxygen Electrode Arrays for Imaging and Multiplexed Measurements of the Respiratory Activity of Cells
title_fullStr	Bipolar Clark-Type Oxygen Electrode Arrays for Imaging and Multiplexed Measurements of the Respiratory Activity of Cells
title_full_unstemmed	Bipolar Clark-Type Oxygen Electrode Arrays for Imaging and Multiplexed Measurements of the Respiratory Activity of Cells
title_sort	bipolar clark-type oxygen electrode arrays for imaging and multiplexed measurements of the respiratory activity of cells
publisher	American Chemical Society
publishDate	2024
url	http://eprints.um.edu.my/45601/ https://doi.org/10.1021/acsomega.3c09802
_version_	1814933279341543424
score	13.211869

Bipolar Clark-Type Oxygen Electrode Arrays for Imaging and Multiplexed Measurements of the Respiratory Activity of Cells

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