1. Nanogap Dielectric Spectroscopy for Aptamer-Based Protein Detection
Manu Sebastian Mannoor, Teena James, Dentcho V. Ivanov, Les Beadling, William Braunlin 
Biophysical Journal 
Volume 98, Issue 4, Pages (February 2010)
DOI: /j.bpj
Copyright © 2010 Biophysical Society Terms and Conditions

2. Figure 1
Calculated distribution of the electric potential between capacitive electrodes when the position of the second electrode is varied along the x axis with respect to the first electrode. The first electrode is fixed at the position of the y axis (x = 0) and the second electrode is moved away from the first electrode along the x axis. The potential distributions for the positions of the second electrode at x = DL, x = 2 DL, x = 5 DL, and x = 10 DL are shown, where DL = 1/κ. As illustrated in the figure, the potential drop across the double layers is minimized when the separation between the electrodes approaches the width of the electrical double layers (Debye length, 1/κ). This effect reduces the contribution of the double layer impedance to the measured impedance and hence eliminates the so called “electrode polarization effect” in the low frequency dielectric spectrum.
Biophysical Journal  , DOI: ( /j.bpj )
Copyright © 2010 Biophysical Society Terms and Conditions

3. Figure 2
(A) Schematics of the fabrication process flow: (a) photo resist spacers are patterned; (b) gold electrodes formed by sacrificial method; (c and d) deposition and patterning of SiO2 for nanometer spacing; (e) deposition of gold; (f) SiO2 spacer removal; (g) a nanogap capacitive sensing structure. (B) Electron microscopic image of the Au electrodes with nanoscale separation.
Biophysical Journal  , DOI: ( /j.bpj )
Copyright © 2010 Biophysical Society Terms and Conditions

4. Figure 3
Dielectric spectrum of air (dashed) and DI water (solid) between the capacitive electrodes. The similarity in shape of the dielectric spectra of the DI water in the nanogap element to that of ice in a conventional capacitive element (29) suggests the ordering of the water molecules in the nanoscale confinement zone.
Biophysical Journal  , DOI: ( /j.bpj )
Copyright © 2010 Biophysical Society Terms and Conditions

5. Figure 4
Relative permittivity as a function of frequency for various concentrations of buffer solutions obtained (A) using the nanogap capacitive element, and (B) using an interdigitated capacitive element with a gap size of 100 μm. The buffers used were dilutions of SSC buffer, 0.15 M NaCl, 0.015 M sodium citrate, buffered to pH 6.8. Plots (a) and (b) are for air and distilled water, respectively. The remaining plots are dilutions of SSC buffer: (c) 0.05× SSC, (d) 0.1× SSC, (e) 0.25× SSC, (f) 0.5× SSC, and (g) 1× SSC, with ionic strengths of M, M, M, 0.114 M, and M, respectively. All impedance measurements were carried out using an SR785 impedance analyzer. The high values of relative permittivities observed at low frequencies for the interdigitated element in B are attributed to the effect of electrode polarization. These relative permittivities are markedly reduced for the nanogap electrodes in A.
Biophysical Journal  , DOI: ( /j.bpj )
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6. Figure 5
Dashed line shows the dielectric spectrum of a bare nanogap in the presence of 0.5× SSC buffer. The dotted curve shows the spectrum of the same nanogap after coating with a SAM of 100:1 mercaptohexanol/thio-derivatized aptamer. The solid curve shows the nanogap after SAM formation followed by α-thrombin binding. All the measurements were taken in 0.5× SSC buffer solution. The replacement of water molecules (ɛ = 80) by a monolayer of lower dielectric permittivity is reflected as a decrease in the overall sensor permittivity. The protein concentration in these experiments was 1 μM in 0.5× SSC buffer solution.
Biophysical Journal  , DOI: ( /j.bpj )
Copyright © 2010 Biophysical Society Terms and Conditions

7. Figure 6
Dielectric spectrum of the interaction of (1 μM) lysozyme (solid) with the immobilized α-thrombin aptamer (dotted). The measurements were taken in 0.5× SSC buffer solution (dashed). The slight shift in the relative permittivity value can be explained by the nonspecific electrostatic binding between the positively charged protein molecules and the negatively charged aptamer sequences.
Biophysical Journal  , DOI: ( /j.bpj )
Copyright © 2010 Biophysical Society Terms and Conditions

8. Figure 7
Dielectric spectra of a nanogap coated with a SAM containing immobilized control oligomer (incapable of forming the active G-quadruplex structure) in the absence (dotted curve) and presence (solid curve) of α-thrombin. The measurements were taken in 0.5× SSC buffer solution. The protein concentration was 1 μM. The nonspecific electrostatic binding between the positively charged protein molecules and the negatively charged oligonucleotide sequences can be observed as the slight shift in the relative permittivity. The dashed curve shows the permittivity of a solution of 0.5× SSC with no oligomer or protein present.
Biophysical Journal  , DOI: ( /j.bpj )
Copyright © 2010 Biophysical Society Terms and Conditions