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Investigating Flagella-Driven Motility in Escherichia coli by Applying Three Established Techniques in a Series
Published on: May 10, 2020
Controlling the shape of filamentous cells of Escherichia coli
Shoji Takeuchi1, Willow R DiLuzio, Douglas B Weibel
1Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, Massachusetts 02138, USA.
Abstract:
This paper describes a technique for growing filamentous cells of Escherichia coli with defined shapes, including crescents, zigzags, sinusoids, and spirals. The procedure begins with the fabrication of embossed microchambers in agarose. Cells are trapped in the chambers by placing a flat, flexible "ceiling", either a slab of agarose or poly(dimethylsiloxane), against an agarose mold on which a suspension of cells has been added; the use of agarose keeps cells hydrated and allows nutrients to diffuse into the chambers. Cells grown in microchambers in the presence of cephalexin grow into a multinucleate, nonseptate, filamentous phenotype and adopt the shape of the microchambers. The resulting cells are motile and can be released by removing the "ceiling" from the agarose microchambers and rinsing the cells into solution.
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