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Cultivation and Imaging of S. latissima Embryo Monolayered Cell Sheets Inside Microfluidic Devices

The culturing and investigation of individual marine specimens in lab environments is crucial to further our understanding of this highly complex ecosystem. However, the obtained results and their relevance are often limited by a lack of suitable experimental setups enabling controlled specimen grow...

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Autores principales: Clerc, Thomas, Boscq, Samuel, Attia, Rafaele, Kaminski Schierle, Gabriele S., Charrier, Bénédicte, Läubli, Nino F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9687954/
https://www.ncbi.nlm.nih.gov/pubmed/36421119
http://dx.doi.org/10.3390/bioengineering9110718
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author Clerc, Thomas
Boscq, Samuel
Attia, Rafaele
Kaminski Schierle, Gabriele S.
Charrier, Bénédicte
Läubli, Nino F.
author_facet Clerc, Thomas
Boscq, Samuel
Attia, Rafaele
Kaminski Schierle, Gabriele S.
Charrier, Bénédicte
Läubli, Nino F.
author_sort Clerc, Thomas
collection PubMed
description The culturing and investigation of individual marine specimens in lab environments is crucial to further our understanding of this highly complex ecosystem. However, the obtained results and their relevance are often limited by a lack of suitable experimental setups enabling controlled specimen growth in a natural environment while allowing for precise monitoring and in-depth observations. In this work, we explore the viability of a microfluidic device for the investigation of the growth of the alga Saccharina latissima to enable high-resolution imaging by confining the samples, which usually grow in 3D, to a single 2D plane. We evaluate the specimen’s health based on various factors such as its growth rate, cell shape, and major developmental steps with regard to the device’s operating parameters and flow conditions before demonstrating its compatibility with state-of-the-art microscopy imaging technologies such as the skeletonisation of the specimen through calcofluor white-based vital staining of its cell contours as well as the immunolocalisation of the specimen’s cell wall. Furthermore, by making use of the on-chip characterisation capabilities, we investigate the influence of altered environmental illuminations on the embryonic development using blue and red light. Finally, live tracking of fluorescent microspheres deposited on the surface of the embryo permits the quantitative characterisation of growth at various locations of the organism.
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spelling pubmed-96879542022-11-25 Cultivation and Imaging of S. latissima Embryo Monolayered Cell Sheets Inside Microfluidic Devices Clerc, Thomas Boscq, Samuel Attia, Rafaele Kaminski Schierle, Gabriele S. Charrier, Bénédicte Läubli, Nino F. Bioengineering (Basel) Article The culturing and investigation of individual marine specimens in lab environments is crucial to further our understanding of this highly complex ecosystem. However, the obtained results and their relevance are often limited by a lack of suitable experimental setups enabling controlled specimen growth in a natural environment while allowing for precise monitoring and in-depth observations. In this work, we explore the viability of a microfluidic device for the investigation of the growth of the alga Saccharina latissima to enable high-resolution imaging by confining the samples, which usually grow in 3D, to a single 2D plane. We evaluate the specimen’s health based on various factors such as its growth rate, cell shape, and major developmental steps with regard to the device’s operating parameters and flow conditions before demonstrating its compatibility with state-of-the-art microscopy imaging technologies such as the skeletonisation of the specimen through calcofluor white-based vital staining of its cell contours as well as the immunolocalisation of the specimen’s cell wall. Furthermore, by making use of the on-chip characterisation capabilities, we investigate the influence of altered environmental illuminations on the embryonic development using blue and red light. Finally, live tracking of fluorescent microspheres deposited on the surface of the embryo permits the quantitative characterisation of growth at various locations of the organism. MDPI 2022-11-21 /pmc/articles/PMC9687954/ /pubmed/36421119 http://dx.doi.org/10.3390/bioengineering9110718 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Clerc, Thomas
Boscq, Samuel
Attia, Rafaele
Kaminski Schierle, Gabriele S.
Charrier, Bénédicte
Läubli, Nino F.
Cultivation and Imaging of S. latissima Embryo Monolayered Cell Sheets Inside Microfluidic Devices
title Cultivation and Imaging of S. latissima Embryo Monolayered Cell Sheets Inside Microfluidic Devices
title_full Cultivation and Imaging of S. latissima Embryo Monolayered Cell Sheets Inside Microfluidic Devices
title_fullStr Cultivation and Imaging of S. latissima Embryo Monolayered Cell Sheets Inside Microfluidic Devices
title_full_unstemmed Cultivation and Imaging of S. latissima Embryo Monolayered Cell Sheets Inside Microfluidic Devices
title_short Cultivation and Imaging of S. latissima Embryo Monolayered Cell Sheets Inside Microfluidic Devices
title_sort cultivation and imaging of s. latissima embryo monolayered cell sheets inside microfluidic devices
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9687954/
https://www.ncbi.nlm.nih.gov/pubmed/36421119
http://dx.doi.org/10.3390/bioengineering9110718
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