Chapter 4: Results and Discussion
4.3 Immobilization of Biomolecules onto the Gold-Silicide Nanowire and Electrical Properties
4.3.6 The immobilization of mouse-IgG and anti-mouse IgG onto gold-silicide nanowires
4.3.6 The immobilization of mouse-IgG and anti-mouse IgG onto gold-silicide nanowires
We choose the process of annealing by furnace at 500℃ for this experiment according to the previous results of the measurements of conductance and fluorescent images. The samples were immobilized with mouse-IgG and FITC-conjugated anti-mouse IgG, and then directly Figure 4.35: The conductance of the gold-silicide nanowires of various widths with
biotin and streptavidin immobilization.
observed under the fluorescent microscope. The SEM morphology in Figure 4.36 showed the pad and various nanowires of gold-silicide. The nanowires were fabricated as our expected widths. The binding of the mouse-IgG and anti-mouse IgG onto the nanowires and pads were too tiny to be seen in the images.
(a) (b)
(c) (d)
(e)
Figure 4.36: The SEM images of the gold-silicide nanowires with various widths annealing at 500℃ by furnace then immobilizing mouse-IgG and hybridizing FITC-conjugated anti-mouse IgG. (a) 80 nm (b) 100 nm (c) 120 nm (d) 200 nm (e) pad.
Figure 4.37 revealed the fluorescent images of the samples. The upper image was the sample modified by 1,2-ethanedithiol and sulfoSMCC and successful immobilized
mouse-IgG and FITC-conjugated anti-mouse IgG. The lower image was the sample without modification can not immobilize mouse-IgG and FITC-conjugated anti-mouse IgG.
The electrical properties of the samples were shown in Figure 4.38. The samples with linker modification could immobilize mouse-IgG and have no obvious changes (Figure 4.38a). When the anti-mouse IgG added to combine with mouse-IgG, the conductance Figure 4.37: The images of the gold-silicide pads and nanowires annealing at 500℃
by furnace. The fluorescence of FITC-conjugated anti-mouse IgG with (upper) and without (lower) surface modification.
increased significantly (Figure 4.38b). If we wash the sample with boiling water for 10 minutes, the mouse-IgG and anti-mouse IgG will denature and can not bind to each other [10].
The conductance restores to the value before immobilization. The sample without surface modification in Figure 4.38c could evaluate the non-specific binding of the mouse-IgG and alter some changes for the electrical property. The non-specific binding of mouse-IgG could fix the anti-mouse IgG. The simultaneous existence of mouse-IgG and anti-mouse IgG on the surface of the gold-silicide nanowire could affect the electrical property.
(a) (c)
(b)
Figure 4.38: The conductance of the gold-silicide nanowires of various widths with IgG and anti-IgG immobilization.
Although the BSA and the detergent Tween 20 were already used in the experiment, we still need some efforts to reduce the non-specific binding in the future. In a short conclusion, the gold-silicide nanowire could use to immobilize a specific biomolecule of interest by our surface modification and binding techniques. We could direct observe the specific binding under fluorescent microscope or measure the changes of electrical properties. The change maybe due to that the current flows extremely close to the surface, the biological
macromolecules bound to the surface of a nanowire and undergoing a binding event with conformational change or change of charge state, may thus affect the current flow in the nanowire. This mechanism is beneficial for a biosensor with state-of-the-art semiconductor manufacturing to detect low concentration of biomolecules.
Chapter 5: Conclusions
The immobilization between the solid support and biomolecules is very important for the interdisciplinary studies of biotechnology and semiconductor fields. The key to
immobilization is the surface modification. We firstly immobilize rhodamine by APTES and glutaraldehyde modification onto various substrates and find out the best solid support is silicon dioxide. The active enzyme, namely sulfotransferase, is also successfully immobilized by APTES and sulfoSMCC modification on the silicon dioxide and still has the activity. We also propose a new material of gold-silicide nanowires to immobilize biomolecules. We fabricate the gold-silicide nanowires by annealing gold-coated poly-silicon nanowires, followed with gold removal by aqua regia. The gold-silicide nanowires have controllable widths and lengths. After surface modification with 1,2-ethanedithiol and SulfoSMCC, the gold-silicide nanowires readily immobilize biomolecules such as hydrazide-biotin and IgG.
The biotin and IgG could bind to streptavidin and anti-IgG, respectively. We observe the specific biomolecules under fluorescent microscope and measure the changes of electrical property. The proposed gold-silicide nanowires, for the first time, can be a very effective means to sense various important biomolecules. These devices also possess the advantages of very high sensitivity, label free and low cost.
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