Thursday, March 28, 2013

Differential interferometric particle tracking on the subnanometer- and submillisecond-scale

Dennis Müller, Dieter R. Klopfenstein, and Rainer G. Ulbrich

We describe an interferometric method to measure the movement of a subwavelength probe particle relative to an immobilized reference particle with high spatial (Δx = 0.9nm) and temporal (Δt = 200μs) resolution. The differential method eliminates microscope stage drift. An upright microscope is equipped with laser dark field illumination (λ0 = 532nm, P0 = 30mW) and a compact modified Mach-Zehnder interferometer is mounted on the camera exit of the microscope, where the beams of scattered light of both particles are combined. The resulting interferograms provide in two channels subnanometer information about the motion of the probe particle relative to the reference particle. The interferograms are probed with two avalanche photodiodes. We applied this method to measuring the movement of kinesin along microtubules and were able to resolve the generic 8-nm steps at high ATP concentrations without external forces.
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