A sol or colloidal dispersion of gold nanoparticles in a solution, generally freshwater, is known as colloidal gold. For spherical particles smaller than 100 nm, the colloid is typically an intense red color, whereas, for larger spherical particles or nanorods, the colloid is blue/purple. Gold nanoparticles are the topic of extensive investigation due to their optical, electrical, and molecular-recognition characteristics, with numerous prospective or promised uses in a wide range of fields, including Electronics, nanotechnology, materials science, and healthcare are among areas in which electron microscopy use.
Because of the nanoparticle’s interactions with visible light, artists have employed colloidal gold for millennia. Depending on particle size, shape, local refractive index, and aggregation state, gold nanoparticles absorb and scatter light, resulting in colors ranging from bright reds to blues to black, and eventually clear and colorless. Because of a phenomenon known as localized surface plasmon resonance, which occurs when conduction electrons on the nanoparticle’s surface vibrate in resonance with incoming light, these hues appear.
Effect of Size
As the size of the nanoparticle rises, the wavelength of light absorbed increases as well. Pseudo-spherical gold nanoparticles with diameters of less than 30 nm, for example, exhibit a peak LSPR absorption of 530 nm.
Effect of Local Refractive Index
The environment in which colloidal gold suspend can also affect the apparent hue of a gold nanoparticle solution. Because the refractive index at the nanoparticle surface determines the optical characteristics of gold nanoparticles, both molecules directly adhering to the nanoparticle surface and the nanoparticle solvent may impact observable visual features.
Effect of Aggregation
The optical characteristics of gold nanoparticles vary as they aggregate because the adequate particle size, shape, and dielectric environment change.
Methods and Materials
All appropriate international, national, and institutional standards for the care and use of animals were followed to minimize suffering. The Shandong Committee on Animal Ethics authorized the animal operations (permit number: 201750017). Two young adult BALB/c mice (35 days old) were utilized in the vaccination protocols. They were kept at Shandong Agricultural University with free access to water and food, relative humidity of 40–60 percent, and a 12–12 h light-dark cycle. The rapid cervical dislocation use to euthanize mice.
Reagents and Materials
Our laboratory identified and maintained goose parvovirus, avian influenza viruses (H5, H7, and H9), duck hepatitis virus, nembutsu virus, fowl adenovirus, goose reovirus, and muscovy duck parvovirus. Our team used sucrose density gradient centrifugation to separate the pure GPV from the GPV-positive allantoic fluid. In earlier research, recombinant VP3 protein produce using a prokaryotic expression system, and the BCA Protein Assay Kit determined VP3 concentration to be 3.34 mg/ml. From Boster Biological Technology Co., Ltd, we obtained goat anti-mouse IgG antibody and bovine serum albumin (BSA) (Wuhan, China).
Preparation of Monoclonal Antibodies
Anti-GPV monoclonal antibodies (anti-GPV Mab) were produced with various modifications described by Yin et al. (2010). In a nutshell, pure GPV suspension was verified by PCR and SDS-PAGE, then emulsified in a 1:1 ratio with Freund’s adjuvant and vaccinated BALB/c mice with it. The fluid at a volume of 0.2 ml. Fusion of spleen cells with SP2/0 myeloma cells produced hybridomas. Indirect ELISA use to assess the reactivity and specificity of hybridoma cells to GPV.
Preparation of Polyclonal Antibody
According to a previously published study, polyclonal antiserum against GPV produce with minor changes (Ma et al., 2017). It produce by immunizing rabbits with recombinant VP3 protein and then coating them with a coating reagent on an NC membrane (test line). The following are the specific steps: Two healthy rabbits inject into the back by various sites subcutaneous injection with 200 g VP3 protein, mixed and emulsified with the same amount of FCA or FIA. The whole vaccination treatment consisted of four shots, the first of which was FCA and the second of which was FIA.
Synthesis of Colloidal Gold
Before use, all glassware, including beakers and flasks, wash in ultrapure water, pretreated in aqua regia, then rinsed and dried in ultrapure water. According to a previously published approach, colloidal gold nanoparticles with a mean particle diameter of 20 nm generate by reducing gold chloride with 1 percent sodium citrate (Zhang et al., 2015).
Using electric heating, ultrapure water (100 mL) heat to boiling point in an Erlenmeyer flask with a magnetic stirrer. Then 1 mL of 1% chloroauric acid solution quickly add to the flask, followed by 3.0 mL of 1% sodium citrate solution promptly added to the solution under stirring. The answer heat for another 10 minutes when the color changed from blue to dark red, then stopped heating, agitated for 15 minutes, cooled, and kept at 4°C with 0.05 percent sodium azide.
Preparation of the Colloidal Gold–Mab Conjugate
Initially, the optimal Mab concentration for conjugation with a colloidal gold solution establish (Liu et al., 2012). The Mab dilute to different concentrations in PBS (0.015 M, pH 7.4) (0 g/ml, 5 g/ml, 7.5 g/ml, 10 g/ml, 20 g/ml, 30 g/ml, 50 g/ml), and 20 l of each concentration add to 1 ml of colloidal gold solution (pH 8.5) with mild shaking for 10 minutes. After that, each tube received 100 l of 10% NaCl, which agitate for another 10 minutes.
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