Tuesday, January 28, 2014

Optical tweezers assisted imaging of the Z-ring in Escherichia coli: measuring its radial width

G Carmon, P Kumar and M Feingold

Using single-beam, oscillating optical tweezers we can trap and rotate rod-shaped bacterial cells with respect to the optical axis. This technique allows imaging fluorescently labeled three-dimensional sub-cellular structures from different, optimized viewpoints. To illustrate our method we measure D, the radial width of the Z-ring in unconstricted Escherichia coli. We use cells that express FtsZ-GFP and have their cytoplasmic membrane stained with FM4-64. In a vertically oriented cell, both the Z-ring and the cytoplasmic membrane images appear as symmetric circular structures that lend themselves to quantitative analysis. We found that D cong 100 nm, much larger than expected.

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