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Polarization-dependent fluorescence of proteins bound to nanopore-confined lipid bilayers
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10.1063/1.2972143
/content/aip/journal/jcp/129/9/10.1063/1.2972143
http://aip.metastore.ingenta.com/content/aip/journal/jcp/129/9/10.1063/1.2972143
View: Figures

Figures

Image of FIG. 1.
FIG. 1.

SEM image of commercial AAO after depositing streptavidin bound to biotinilated lipid bilayers. The pores are extending throughout the sample (approximately thick).

Image of FIG. 2.
FIG. 2.

(a) System configuration. (b) Fluorescence spectrum of DTAF-streptavidin bound to biotinilated lipid bilayers in AAO. (c) Fluorescence peak intensity as a function of time (filled diamonds) and least squares fit to (solid line). (d) Fluorescence spectra of biotinitaled lipid bilayers deposited into either exposed or Au coated AAO substrates before (filled diamonds) and after being introduced to the streptavidin. The samples were not exposed to the laser light prior to taking the data hence the relative large signal.

Image of FIG. 3.
FIG. 3.

(a) Experimental configuration with marked - and -polarization modes. (b) Fluorescence peak intensity for -polarized incident beam. Normal incidence position is at . (c) Repeated experiment for exposed (uncoated) AAO substrate (1) after one month: the curve has the same extinction ratio , albeit with an overall intensity value differences. The inset indicates out-of-plane tilt. (d) The sample was rotated azimuthally (in-plane rotation) by 90° and the experiments were repeated.

Image of FIG. 4.
FIG. 4.

(a) Fluorescence peak intensity for -polarized incident beam. Normal incidence position is at with the exception of for the exposed sample (1). (b) The samples were azimuthally rotated by 90° and the tilt experiments were repeated. Normal incidence position is at with the exception of for the exposed samples.

Image of FIG. 5.
FIG. 5.

(a) Fluorescence peak intensity as a function of azimuthal angle (in-plane rotations as indicated by the inset) for the exposed (uncoated) sample (2) at normal incidence. (b) Radial plot: the asymmetry in the curve is likely due to sample inhomogeneities. (c) Fluorescence as a function of azimuthal angle for Au-coated sample (2) at normal incidence. The purple curve is a shifted sinusoidal curve indicating a threefold symmetry. (d) Repeated measurements demonstrate acceptable reproducibility for the radial plot.

Image of FIG. 6.
FIG. 6.

Simulations: normalized fluorescence signal for -pol (red line) and -pol (blue dash line) modes. The molecules are assumed to be aligned along the pore axis but the position below AAO surface is insignificant.

Image of FIG. 7.
FIG. 7.

Unit vectors and ideal arrangement of holes.

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/content/aip/journal/jcp/129/9/10.1063/1.2972143
2008-09-02
2014-04-24
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752b84549af89a08dbdd7fdb8b9568b5 journal.articlezxybnytfddd
Scitation: Polarization-dependent fluorescence of proteins bound to nanopore-confined lipid bilayers
http://aip.metastore.ingenta.com/content/aip/journal/jcp/129/9/10.1063/1.2972143
10.1063/1.2972143
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