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Single molecule tracking scheme using a near-field scanning optical microscope

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4 Author(s)
Decca, R.S. ; Department of Physics, Indiana University Purdue University Indianapolis, 402 N. Blackford Street, Indianapolis, Indiana 46202 ; Lee, C.-W. ; Lall, S. ; Wassall, S.R.

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The use of a near-field scanning optical microscope (NSOM) to measure the diffusion of fluorescently tagged molecules on a quasi-two-dimensional system is described. The heart of the apparatus lies in the circular motion of the NSOM probe, allowing the detection of small displacements on single molecules. Using a 100 nm tip, and a simulated diffusion coefficient of D∼1×10-12m2/s, we found that molecules can be tracked within ∼40 nm over several seconds. We foresee that D as large as ∼5×10-12m2/s can be measured with minor modifications to the experimental setup. These values of D encompass those found in many liquid crystalline systems, particularly lipids in bilayer membranes. © 2002 American Institute of Physics.

Published in:

Review of Scientific Instruments  (Volume:73 ,  Issue: 7 )