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Nanodroplet sampling processing platform for amine containing metabolites in single Yarrowia lipolytical fungal protoplasts


EMSL Project ID
60842

Abstract

Single-cell metabolomics of bioproduction organisms such as Yarrowia lipolytica could significantly enhance our understanding of functional heterogeneity, yet remains a challenge due to limited input material, low retention on liquid-chromatography (LC) columns, and poor mass spectrometry (MS) ionization efficiency. To address these challenges, we propose using the cellenONE single-cell isolation system in combination with nanoPOTS and tandem mass tags (TMT) to create a state-of-the-art single-cell LC-MS metabolomics workflow. The cellenONE will enable facile isolation of these smaller (5-8 µm) single-cells while nanoPOTS will minimize surface adsorption and sample loss. To further resolve trace sample size issues, amine-containing metabolites will be reacted with TMT to improve LC retention, ionization efficiency, and boost-signal via a TMT reference channel. We aim to enhance EMSL’s metabolomics capability and extend its application to high-throughput single-cell metabolomic profiling of fungal and plant cells.

Project Details

Start Date
2023-04-17
End Date
N/A
Status
Active

Team

Principal Investigator

Sarah Williams
Institution
Environmental Molecular Sciences Laboratory

Team Members

James Fulcher
Institution
Environmental Molecular Sciences Laboratory

Aivett Bilbao Pena
Institution
Environmental Molecular Sciences Laboratory

Nathalie Munoz Munoz
Institution
Environmental Molecular Sciences Laboratory

Erin Bredeweg
Institution
Environmental Molecular Sciences Laboratory