title: Laser capture microdissection in Ectocarpus siliculosus: the pathway to cell-specific transcriptomics in brown algae.
abstract: Laser capture microdissection (LCM) facilitates the isolation of individual cells from tissue sections, and when combined with RNA amplification techniques, it is an extremely powerful tool for examining genome-wide expression profiles in specific cell-types. LCM has been widely used to address various biological questions in both animal and plant systems, however, no attempt has been made so far to transfer LCM technology to macroalgae. Macroalgae are a collection of widespread eukaryotes living in fresh and marine water. In line with the collective effort to promote molecular investigations of macroalgal biology, here we demonstrate the feasibility of using LCM and cell-specific transcriptomics to study development of the brown alga Ectocarpus siliculosus. We describe a workflow comprising cultivation and fixation of algae on glass slides, laser microdissection, and RNA amplification. To illustrate the effectiveness of the procedure, we show qPCR data and metrics obtained from cell-specific transcriptomes generated from both upright and prostrate filaments of Ectocarpus.
authors.affiliation: Department of Plant Sciences, University of Oxford Oxford, UK.
authors.forename: Denis
authors.initials: D
authors.lastname: Saint-Marcoux
authors.affiliation: CNRS, Sorbonne Université, UPMC Univ Paris 06, UMR 8227, Integrative Biology of Marine Models, Station Biologique de Roscoff Roscoff, France.
authors.forename: Bernard
authors.initials: B
authors.lastname: Billoud
authors.affiliation: Department of Plant Sciences, University of Oxford Oxford, UK.
authors.forename: Jane A
authors.initials: JA
authors.lastname: Langdale
authors.affiliation: CNRS, Sorbonne Université, UPMC Univ Paris 06, UMR 8227, Integrative Biology of Marine Models, Station Biologique de Roscoff Roscoff, France.
authors.forename: Bénédicte
authors.initials: B
authors.lastname: Charrier
keywords: brown algae; cell differentiation; cell-specific transcriptomics; laser capture microdissection; seaweed