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Results: 5

1.
Figure 2

Figure 2. From: Nanochannel Based Single Molecule Recycling.

Single Molecule Recycling of dsDNA13mers. Single dsDNA13mers are captured with the single molecule recycler and measured at the seconds time scale. Donor curves are plotted in green and acceptor curves are plotted in red. For clarity, acceptor time traces are plotted towards negative values. A: Recycling of the same dsDNA13mer 144 times in ~3 s with a 6 ms delay in reversal of electroosmosis after the molecule exits the focus. B: Same data as in panel A, plotted with an expanded time scale demonstrating the optical shutter effect.

John F. Lesoine, et al. Nano Lett. ;12(6):3273-3278.
2.
Figure 5

Figure 5. From: Nanochannel Based Single Molecule Recycling.

Repeated FRET measurements on a single OxlT membrane protein. Panel A: Fluorescence time trace of donor (green) and acceptor (red). Panel B: Corresponding energy transfer efficiency (ETE). The mean ETE is 0.315 (solid horizontal line) and the standard deviation is 0.086 (dashed horizontal lines). The variations in ETE are the result of shot noise. The inset shows the homology model of OxlT with the donor and acceptor sites (red features).

John F. Lesoine, et al. Nano Lett. ;12(6):3273-3278.
3.
Figure 4

Figure 4. From: Nanochannel Based Single Molecule Recycling.

Single Molecule Recycling combined with spFRET. Distributions of energy transfer efficiency (ETE) for dsDNA molecules of two different lengths (13 base pairs and 15 base pairs). Panel B: Distributions resulting from measuring different dsDNA only once (standard diffusion-based experiment). 65030 transits are used for the dsDNA13mer distribution, 120435 transits for the dsDNA15mer distribution, and 148830 transits for the mixture. Panel B: Same as in Panel A but sampling each dsDNA molecule at least nine times. Single molecule recycling improves the resolution and accuracy. 10789 transits are used for the dsDNA13mer distribution, 22875 transits for the dsDNA15mer distribution, and 4341 transits for the mixture. The distribution were created using the function, ksdensity, in MATLAB with a width of (A) 0.015 and (B–D) 0.02.

John F. Lesoine, et al. Nano Lett. ;12(6):3273-3278.
4.
Figure 1

Figure 1. From: Nanochannel Based Single Molecule Recycling.

Schematic of Single Molecule Recycling. Donor-acceptor labeled dsDNA are transported along a nanofluidic channel (cross-section ~ 600nm × 400nm) by use of electroosmotic flow. A controller reverses the flow in the nanochannel after detecting a fluorescence burst, causing the molecule to pass the excitation focus again. This procedure is repeated and makes it possible to sample the same molecule over and over again at periodic times that can be adjusted by the flow speed and the parameters of the flow controller. BS: Beam splitter; F: Filter; NA: Numerical aperture. Inset (top left): SEM image of fabricated nanochannels.

John F. Lesoine, et al. Nano Lett. ;12(6):3273-3278.
5.
Figure 3

Figure 3. From: Nanochannel Based Single Molecule Recycling.

Time Between Consecutive Passes. Distribution of times between one molecule exiting the focus and the next consecutive molecule entering the focus. The peaks at 36.1 ± .1 ms and 65.43 ± .06 ms with respective standard deviations 5.2 ± 0.2 ms and 8.2 ± 0.1 ms correspond to twice the turnaround times of 15 s and 30 s plus twice the software loop time ≈ 2.9 ms for experiments with dsDNA15mers (circles) and dsDNA13mers (triangles) respectively. The widths of the Gaussians are the result of the uncertainty in position due to diffusion. The Gaussians are superimposed on exponential distributions, which describe the times between arrivals of different molecules into the trap.

John F. Lesoine, et al. Nano Lett. ;12(6):3273-3278.

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