Live Single-Cell Metabolomics With Matrix-Free Laser/Desorption Ionization Mass Spectrometry to Address Microalgal Physiology
Unicellular phototrophic algae can form massive blooms with up to millions of individual cells per milliliter in freshwater and marine ecosystems. Despite the temporal dominance of bloom formers many algal species can co-exist and compete for nutrients and space, creating a complex and diverse commu...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2019-02-01
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| Series: | Frontiers in Plant Science |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/article/10.3389/fpls.2019.00172/full |
| _version_ | 1818135165343367168 |
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| author | Tim U. H. Baumeister Marine Vallet Filip Kaftan Aleš Svatoš Georg Pohnert Georg Pohnert |
| author_facet | Tim U. H. Baumeister Marine Vallet Filip Kaftan Aleš Svatoš Georg Pohnert Georg Pohnert |
| author_sort | Tim U. H. Baumeister |
| collection | DOAJ |
| description | Unicellular phototrophic algae can form massive blooms with up to millions of individual cells per milliliter in freshwater and marine ecosystems. Despite the temporal dominance of bloom formers many algal species can co-exist and compete for nutrients and space, creating a complex and diverse community. While microscopy and single cell genomics can address the taxonomic inventory, the cellular metabolome has yet to be thoroughly explored to determine the physiological status of microalgae. This might, however, provide a key to understand the observed species diversity in the homogeneous environment. Here, we introduce an effective, rapid and versatile method to analyze living single cells from aqueous substrata with laser-desorption/ionization mass spectrometry (LDI-MS) using a simple and inexpensive matrix-free support. The cells deposited on a cultivation-medium wetted support are analyzed with minimal disturbance as they remain in their natural viable state until their disruption during LDI-MS. Metabolites desorbed from single cells are analyzed on High-Resolution Mass Spectrometry (HR-MS) using the Orbitrap FT-MS technology to fingerprint cellular chemistry. This live single-cell mass spectrometry (LSC-MS) allows assessing the physiological status and strain-specifics of different microalgae, including marine diatoms and freshwater chlorophytes, at the single-cell level. We further report a reliable and robust data treatment pipeline to perform multivariate statistics on the replicated LSC-MS data. Comparing single cell MS spectra from natural phytoplankton samples and from laboratory strains allows the identification and discrimination of inter and intra-specific metabolic variability and thereby has promising applications in addressing highly complex phytoplankton communities. Notably, the herein described matrix-free live-single-cell LDI-HR-MS approach enables monitoring dynamics of the plankton and might explain why key-players survive, thrive, avoid selective feeding or pathogenic virus and bacteria, while others are overcome and die. |
| first_indexed | 2024-12-11T09:20:10Z |
| format | Article |
| id | doaj.art-8cf18e349ede419c921c773d661e133b |
| institution | Directory Open Access Journal |
| issn | 1664-462X |
| language | English |
| last_indexed | 2024-12-11T09:20:10Z |
| publishDate | 2019-02-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Plant Science |
| spelling | doaj.art-8cf18e349ede419c921c773d661e133b2022-12-22T01:13:16ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2019-02-011010.3389/fpls.2019.00172423908Live Single-Cell Metabolomics With Matrix-Free Laser/Desorption Ionization Mass Spectrometry to Address Microalgal PhysiologyTim U. H. Baumeister0Marine Vallet1Filip Kaftan2Aleš Svatoš3Georg Pohnert4Georg Pohnert5Max Planck Fellow Group on Plankton Community Interaction, Max Planck Institute for Chemical Ecology, Jena, GermanyMax Planck Fellow Group on Plankton Community Interaction, Max Planck Institute for Chemical Ecology, Jena, GermanyResearch Group Mass Spectrometry/Proteomics, Max Planck Institute for Chemical Ecology, Jena, GermanyResearch Group Mass Spectrometry/Proteomics, Max Planck Institute for Chemical Ecology, Jena, GermanyMax Planck Fellow Group on Plankton Community Interaction, Max Planck Institute for Chemical Ecology, Jena, GermanyInstitute for Inorganic and Analytical Chemistry, Bioorganic Analytics, Friedrich Schiller University Jena, Jena, GermanyUnicellular phototrophic algae can form massive blooms with up to millions of individual cells per milliliter in freshwater and marine ecosystems. Despite the temporal dominance of bloom formers many algal species can co-exist and compete for nutrients and space, creating a complex and diverse community. While microscopy and single cell genomics can address the taxonomic inventory, the cellular metabolome has yet to be thoroughly explored to determine the physiological status of microalgae. This might, however, provide a key to understand the observed species diversity in the homogeneous environment. Here, we introduce an effective, rapid and versatile method to analyze living single cells from aqueous substrata with laser-desorption/ionization mass spectrometry (LDI-MS) using a simple and inexpensive matrix-free support. The cells deposited on a cultivation-medium wetted support are analyzed with minimal disturbance as they remain in their natural viable state until their disruption during LDI-MS. Metabolites desorbed from single cells are analyzed on High-Resolution Mass Spectrometry (HR-MS) using the Orbitrap FT-MS technology to fingerprint cellular chemistry. This live single-cell mass spectrometry (LSC-MS) allows assessing the physiological status and strain-specifics of different microalgae, including marine diatoms and freshwater chlorophytes, at the single-cell level. We further report a reliable and robust data treatment pipeline to perform multivariate statistics on the replicated LSC-MS data. Comparing single cell MS spectra from natural phytoplankton samples and from laboratory strains allows the identification and discrimination of inter and intra-specific metabolic variability and thereby has promising applications in addressing highly complex phytoplankton communities. Notably, the herein described matrix-free live-single-cell LDI-HR-MS approach enables monitoring dynamics of the plankton and might explain why key-players survive, thrive, avoid selective feeding or pathogenic virus and bacteria, while others are overcome and die.https://www.frontiersin.org/article/10.3389/fpls.2019.00172/fullmicroalgaesingle-cell metabolomicslive single cell mass spectrometrymatrix-free laser desorption/ionization-mass spectrometrymultivariate statisticsdiatoms Coscinodiscus (Bacillariophyceae) |
| spellingShingle | Tim U. H. Baumeister Marine Vallet Filip Kaftan Aleš Svatoš Georg Pohnert Georg Pohnert Live Single-Cell Metabolomics With Matrix-Free Laser/Desorption Ionization Mass Spectrometry to Address Microalgal Physiology Frontiers in Plant Science microalgae single-cell metabolomics live single cell mass spectrometry matrix-free laser desorption/ionization-mass spectrometry multivariate statistics diatoms Coscinodiscus (Bacillariophyceae) |
| title | Live Single-Cell Metabolomics With Matrix-Free Laser/Desorption Ionization Mass Spectrometry to Address Microalgal Physiology |
| title_full | Live Single-Cell Metabolomics With Matrix-Free Laser/Desorption Ionization Mass Spectrometry to Address Microalgal Physiology |
| title_fullStr | Live Single-Cell Metabolomics With Matrix-Free Laser/Desorption Ionization Mass Spectrometry to Address Microalgal Physiology |
| title_full_unstemmed | Live Single-Cell Metabolomics With Matrix-Free Laser/Desorption Ionization Mass Spectrometry to Address Microalgal Physiology |
| title_short | Live Single-Cell Metabolomics With Matrix-Free Laser/Desorption Ionization Mass Spectrometry to Address Microalgal Physiology |
| title_sort | live single cell metabolomics with matrix free laser desorption ionization mass spectrometry to address microalgal physiology |
| topic | microalgae single-cell metabolomics live single cell mass spectrometry matrix-free laser desorption/ionization-mass spectrometry multivariate statistics diatoms Coscinodiscus (Bacillariophyceae) |
| url | https://www.frontiersin.org/article/10.3389/fpls.2019.00172/full |
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