Abstract
Fluorescently labeled polysaccharides enable the visualization of carbohydrate–bacterial interactions and the quantification of carbohydrate hydrolysis rates in cultures and complex communities. Here, we present the method of generating polysaccharides conjugated to the fluorescent molecule, fluoresceinamine. Further, we describe the protocol of incubating these probes in bacterial cultures and complex environmental microbial communities, visualizing bacterial–probe interactions using fluorescence microscopy, and quantifying these interactions using flow cytometry. Finally, we present a novel approach for the in situ metabolic phenotyping of bacterial cells using fluorescently activated cell sorting coupled with omics-based analysis.
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Arnosti C (2003) Fluorescent derivatization of polysaccharides and carbohydrate-containing biopolymers for measurement of enzyme activities in complex media. J Chromatogr B 793(1):181–191. https://doi.org/10.1016/S1570-0232(03)00375-1
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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
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Reintjes, G., Klassen, L., Abbott, D.W. (2023). Visualization of Carbohydrate Uptake Using Fluorescent Polysaccharides. In: Abbott, D.W., Zandberg, W.F. (eds) Carbohydrate-Protein Interactions. Methods in Molecular Biology, vol 2657. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3151-5_18
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DOI: https://doi.org/10.1007/978-1-0716-3151-5_18
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-3150-8
Online ISBN: 978-1-0716-3151-5
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