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Towards high-throughput parallel imaging and single-cell transcriptomics of microbial eukaryotic plankton
Stockholm University, Faculty of Science, Department of Ecology, Environment and Plant Sciences. Stockholm University, Science for Life Laboratory (SciLifeLab).ORCID iD: 0000-0002-3322-599x
Stockholm University, Faculty of Science, Department of Biochemistry and Biophysics. Stockholm University, Science for Life Laboratory (SciLifeLab).ORCID iD: 0000-0003-3053-9392
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Number of Authors: 92024 (English)In: PLOS ONE, E-ISSN 1932-6203, Vol. 19, no 1, article id e0296672Article in journal (Refereed) Published
Abstract [en]

Single-cell transcriptomics has the potential to provide novel insights into poorly studied microbial eukaryotes. Although several such technologies are available and benchmarked on mammalian cells, few have been tested on protists. Here, we applied a microarray single-cell sequencing (MASC-seq) technology, that generates microscope images of cells in parallel with capturing their transcriptomes, on three species representing important plankton groups with different cell structures; the ciliate Tetrahymena thermophila, the diatom Phaeodactylum tricornutum, and the dinoflagellate Heterocapsa sp. Both the cell fixation and permeabilization steps were adjusted. For the ciliate and dinoflagellate, the number of transcripts of microarray spots with single cells were significantly higher than for background spots, and the overall expression patterns were correlated with that of bulk RNA, while for the much smaller diatom cells, it was not possible to separate single-cell transcripts from background. The MASC-seq method holds promise for investigating "microbial dark matter”, although further optimizations are necessary to increase the signal-to-noise ratio.

Place, publisher, year, edition, pages
2024. Vol. 19, no 1, article id e0296672
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Cell and Molecular Biology
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URN: urn:nbn:se:su:diva-228658DOI: 10.1371/journal.pone.0296672ISI: 001150526800053PubMedID: 38241213Scopus ID: 2-s2.0-85182856467OAI: oai:DiVA.org:su-228658DiVA, id: diva2:1854537
Available from: 2024-04-26 Created: 2024-04-26 Last updated: 2024-04-26Bibliographically approved

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Grujčić, VesnaSundh, JohnFoster, Rachel Ann

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Department of Ecology, Environment and Plant SciencesScience for Life Laboratory (SciLifeLab)Department of Biochemistry and Biophysics
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