Growth and Labelling of Cell Wall Components of the Brown Alga Ectocarpus in Microfluidic Chips

Polydimethylsiloxane (PDMS) chips have proven to be suitable environments for the growth of several filamentous organisms. However, depending on the specimen, the number of investigations concerning their growth and cell differentiation is limited. In this work, we monitored the developmental patter...

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Autores principales:	Bénédicte Charrier, Samuel Boscq, Bradley J. Nelson, Nino F. Läubli
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	microfluidics brown alga tip growth on-chip immunolocalisation Ectocarpus filaments Science Q General. Including nature conservation, geographical distribution QH1-199.5
Acceso en línea:	https://doaj.org/article/91b18602a0ad42509ad93a6bfd3a8f3c
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spelling	oai:doaj.org-article:91b18602a0ad42509ad93a6bfd3a8f3c2021-11-15T04:46:17ZGrowth and Labelling of Cell Wall Components of the Brown Alga Ectocarpus in Microfluidic Chips2296-774510.3389/fmars.2021.745654https://doaj.org/article/91b18602a0ad42509ad93a6bfd3a8f3c2021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fmars.2021.745654/fullhttps://doaj.org/toc/2296-7745Polydimethylsiloxane (PDMS) chips have proven to be suitable environments for the growth of several filamentous organisms. However, depending on the specimen, the number of investigations concerning their growth and cell differentiation is limited. In this work, we monitored the developmental pattern of the brown alga Ectocarpus inside PDMS lab-on-chips. Two main methods of inoculation of the lab-on-chip were tested, i.e., via the direct injection of spores into the chamber as well as through the insertion of sporophyte filaments. The resulting growth rate, growth trajectory, cell differentiation, and cell branching were monitored and quantified for 20 days inside 25 or 40 μm parallel channels under standard light and temperature conditions. With growth rates of 2.8 μm⋅h–1, normal growth trajectories and cell differentiation, as well as branching occurring inside the microfluidic environment, the main development steps were shown to be similar to those observed in non-constrained in vitro conditions. Additionally, the labelling of Ectocarpus cell wall polysaccharides using calcofluor for cellulose detection and immunolocalisation with monoclonal antibodies for alginates showed the expected patterns when compared to open space growth evaluated with either epifluorescence or confocal microscopy. Overall, this article describes the experimental conditions for observing and studying the basic unaltered processes of brown algal growth using microfluidic technology which provides the basis for future biochemical and biological researches.Bénédicte CharrierSamuel BoscqBradley J. NelsonNino F. LäubliNino F. LäubliFrontiers Media S.A.articlemicrofluidicsbrown algatip growthon-chip immunolocalisationEctocarpusfilamentsScienceQGeneral. Including nature conservation, geographical distributionQH1-199.5ENFrontiers in Marine Science, Vol 8 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	microfluidics brown alga tip growth on-chip immunolocalisation Ectocarpus filaments Science Q General. Including nature conservation, geographical distribution QH1-199.5
spellingShingle	microfluidics brown alga tip growth on-chip immunolocalisation Ectocarpus filaments Science Q General. Including nature conservation, geographical distribution QH1-199.5 Bénédicte Charrier Samuel Boscq Bradley J. Nelson Nino F. Läubli Nino F. Läubli Growth and Labelling of Cell Wall Components of the Brown Alga Ectocarpus in Microfluidic Chips
description	Polydimethylsiloxane (PDMS) chips have proven to be suitable environments for the growth of several filamentous organisms. However, depending on the specimen, the number of investigations concerning their growth and cell differentiation is limited. In this work, we monitored the developmental pattern of the brown alga Ectocarpus inside PDMS lab-on-chips. Two main methods of inoculation of the lab-on-chip were tested, i.e., via the direct injection of spores into the chamber as well as through the insertion of sporophyte filaments. The resulting growth rate, growth trajectory, cell differentiation, and cell branching were monitored and quantified for 20 days inside 25 or 40 μm parallel channels under standard light and temperature conditions. With growth rates of 2.8 μm⋅h–1, normal growth trajectories and cell differentiation, as well as branching occurring inside the microfluidic environment, the main development steps were shown to be similar to those observed in non-constrained in vitro conditions. Additionally, the labelling of Ectocarpus cell wall polysaccharides using calcofluor for cellulose detection and immunolocalisation with monoclonal antibodies for alginates showed the expected patterns when compared to open space growth evaluated with either epifluorescence or confocal microscopy. Overall, this article describes the experimental conditions for observing and studying the basic unaltered processes of brown algal growth using microfluidic technology which provides the basis for future biochemical and biological researches.
format	article
author	Bénédicte Charrier Samuel Boscq Bradley J. Nelson Nino F. Läubli Nino F. Läubli
author_facet	Bénédicte Charrier Samuel Boscq Bradley J. Nelson Nino F. Läubli Nino F. Läubli
author_sort	Bénédicte Charrier
title	Growth and Labelling of Cell Wall Components of the Brown Alga Ectocarpus in Microfluidic Chips
title_short	Growth and Labelling of Cell Wall Components of the Brown Alga Ectocarpus in Microfluidic Chips
title_full	Growth and Labelling of Cell Wall Components of the Brown Alga Ectocarpus in Microfluidic Chips
title_fullStr	Growth and Labelling of Cell Wall Components of the Brown Alga Ectocarpus in Microfluidic Chips
title_full_unstemmed	Growth and Labelling of Cell Wall Components of the Brown Alga Ectocarpus in Microfluidic Chips
title_sort	growth and labelling of cell wall components of the brown alga ectocarpus in microfluidic chips
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/91b18602a0ad42509ad93a6bfd3a8f3c
work_keys_str_mv	AT benedictecharrier growthandlabellingofcellwallcomponentsofthebrownalgaectocarpusinmicrofluidicchips AT samuelboscq growthandlabellingofcellwallcomponentsofthebrownalgaectocarpusinmicrofluidicchips AT bradleyjnelson growthandlabellingofcellwallcomponentsofthebrownalgaectocarpusinmicrofluidicchips AT ninoflaubli growthandlabellingofcellwallcomponentsofthebrownalgaectocarpusinmicrofluidicchips AT ninoflaubli growthandlabellingofcellwallcomponentsofthebrownalgaectocarpusinmicrofluidicchips
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Growth and Labelling of Cell Wall Components of the Brown Alga Ectocarpus in Microfluidic Chips

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