Visualization of Carbohydrate Uptake Using Fluorescent Polysaccharides

Citation

Reintjes, G., Klassen, L., Abbott, D.W. (2023). Visualization of Carbohydrate Uptake Using Fluorescent Polysaccharides. Methods in Molecular Biology (Clifton, N.J.), [online] 2657 241-249. http://dx.doi.org/10.1007/978-1-0716-3151-5_18

Résumé en langage clair

Fluorescently labelled polysaccharides (FLAPS) are excellent tools to visualize the interactions between carbohydrates and bacteria. In this chapter, we present the method to produce FLAPS. Further, we describe the protocol for incubating FLAPS with bacteria from pure and complex communities, and how to visualize and quantify these samples using fluorescence microscopy and fluorescence activated cell sorting, respectively. Overall, this chapter presents a relatively inexpensive and fast method to identify bacteria that encode enzymes and pathways to metabolize carbohydrates of interest.

Résumé

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.

Date de publication

2023-01-01

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