Summary: | 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 ≅ 100 nm, much larger than expected.
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