Microstamped Petri dishes for scanning electrochemical microscopy analysis of arrays of microtissues.

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 m...

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Autores principales:	Adithya Sridhar, Hans L de Boer, Albert van den Berg, Séverine Le Gac
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/27a1e9369dbd4975b229bb6b4d68ebc9
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spelling	oai:doaj.org-article:27a1e9369dbd4975b229bb6b4d68ebc92021-11-18T08:25:35ZMicrostamped Petri dishes for scanning electrochemical microscopy analysis of arrays of microtissues.1932-620310.1371/journal.pone.0093618https://doaj.org/article/27a1e9369dbd4975b229bb6b4d68ebc92014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24690887/?tool=EBIhttps://doaj.org/toc/1932-6203While 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.Adithya SridharHans L de BoerAlbert van den BergSéverine Le GacPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 4, p e93618 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Adithya Sridhar Hans L de Boer Albert van den Berg Séverine Le Gac Microstamped Petri dishes for scanning electrochemical microscopy analysis of arrays of microtissues.
description	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.
format	article
author	Adithya Sridhar Hans L de Boer Albert van den Berg Séverine Le Gac
author_facet	Adithya Sridhar Hans L de Boer Albert van den Berg Séverine Le Gac
author_sort	Adithya Sridhar
title	Microstamped Petri dishes for scanning electrochemical microscopy analysis of arrays of microtissues.
title_short	Microstamped Petri dishes for scanning electrochemical microscopy analysis of arrays of microtissues.
title_full	Microstamped Petri dishes for scanning electrochemical microscopy analysis of arrays of microtissues.
title_fullStr	Microstamped Petri dishes for scanning electrochemical microscopy analysis of arrays of microtissues.
title_full_unstemmed	Microstamped Petri dishes for scanning electrochemical microscopy analysis of arrays of microtissues.
title_sort	microstamped petri dishes for scanning electrochemical microscopy analysis of arrays of microtissues.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/27a1e9369dbd4975b229bb6b4d68ebc9
work_keys_str_mv	AT adithyasridhar microstampedpetridishesforscanningelectrochemicalmicroscopyanalysisofarraysofmicrotissues AT hansldeboer microstampedpetridishesforscanningelectrochemicalmicroscopyanalysisofarraysofmicrotissues AT albertvandenberg microstampedpetridishesforscanningelectrochemicalmicroscopyanalysisofarraysofmicrotissues AT severinelegac microstampedpetridishesforscanningelectrochemicalmicroscopyanalysisofarraysofmicrotissues
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Microstamped Petri dishes for scanning electrochemical microscopy analysis of arrays of microtissues.

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