The optical spectra of the prepared intracellular and extracellular BC-capped Ag NPs were recorded by a UV–Vis double-beam spectrophotometer (λ-950, Perkin Elmer, USA). FTIR spectroscopy (8400 S Shimadzu, UK) was used to determine the functional groups of chemical components for both. The intracellular and extracellular BC-capped Ag NPs’ morphologies and size were characterized by a high-resolution TEM with an accelerating voltage of 200 kV (HRTEM, JEOL-2010F, JEOL, Japan) and SEM with accelerating voltage ranges from 0.5 to 30 kV (JSM–6510, JEOL, Japan). The particle diameter distribution histograms were determined using HRTEM images of both intracellular and extracellular Ag NPs and using image-J software76 . XRD charts of the intracellular and extracellular BC-capped Ag NPs were measured by X-Ray diffractometer (XRD, Philips X’Pert Pro MRD with CuKα line of wavelength 0.154 nm), operated at 40 kV along with 20 mA in 2θ range from 10° to 80° with a 2° min–1 scanning velocity. DLS and zeta potential studies were also performed for the prepared samples using (Zetasizer Nano ZN, Malvern Panalytical Ltd, UK) at a pH of 7 and a temperature of 25 °C.
Zetasizer nano zn
Manufactured by Malvern Panalytical
Sourced in United Kingdom
The Zetasizer Nano ZN is a dynamic light scattering (DLS) instrument used for the measurement of particle size, zeta potential, and molecular weight. It is capable of analyzing samples with particle sizes ranging from 0.3 nanometers to 10 micrometers.
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9 protocols using zetasizer nano zn
1
Characterization of Biocapped Silver Nanoparticles
2
Nanoparticle Characterization by DLS
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The prepared particles were analyzed for their particle size and size distribution in terms of the average volume diameters and polydispersity index by photon correlation spectroscopy using the particle size analyzer Dynamic Light Scattering (DLS) (Zetasizer Nano ZN, Malvern Panalytical Ltd., Grovewood Road, Malvern, UK) at a fixed angle of 173 °C at 25 °C. Samples were analyzed in triplicate. DLS measures the Brownian motion of nano-sized droplets and relates this movement to an equivalent hydrodynamic diameter (nm). Average droplet size, size distribution curves, and polydispersity index were used to characterize oil droplet dispersion in nanoemulsions. The electrophoretic mobility of oil droplets also reported as z-potential, was measured by the same equipment mentioned above. It determines the surface electrical charge of the droplets dispersed in the continuous phase [22 (link)].
3
Nanoparticle Characterization Protocol
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Particle size, homogenousity of distribution and zeta potentials of nanoparticles were determined using Zeta sizer Nano ZN (Malvern Panalytical Ltd, United Kingdom). Before the measurements, dilution of an aliquot of nanoparticles with ultra purified water, then sonication for 15 min was performed.
4
Nanoparticle Size and Charge Analysis
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The prepared NPs were analyzed for their particle size and size distribution in terms of the average volume diameters and polydispersity index (PDI) by photon correlation spectroscopy using the particle size analyzer Dynamic Light Scattering (DLS) (Zetasizer Nano ZN, Malvern Panalytical Ltd, UK) at a fixed angle of 173°. Samples were analyzed in triplicates the same equipment measured zeta potential to detect the repulsion between NPs21 .
5
HPLC Analysis of Nanoparticles
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The HPLC system (Nouryon, Bohus, Sweden) was equipped with a C18-bonded silica column and silica-based monomeric-type reversed-phase (RP) packing material (Kromasil, C18, 100 Å pore size, 150 × 4.6 mm I.D., 5 µm particle size). The sample was dissolved in a portion of the mobile phase, at 98:2%, including 0.1% trifluoroacetic acid (TFA) in water/acetonitrile, methyl cyanide (MeCN), using five different concentrations. The flow rate was 1 mL/min and we used a 260 nm UV detector. The particle size and zeta potential of the nanoparticles were also tested by dynamic light scattering (DLS) (Zetasizer Nano ZN, Malvern Panalytical Ltd., Malvern, UK) [10 (link),20 (link)].
6
Particle Size and Zeta Potential Analysis
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Samples of SLN, SSN and USN were analyzed for their particle size in terms of the average volume diameters by photon correlation spectroscopy using particle size analyzer Dynamic Light Scattering (DLS) (Zetasizer Nano ZN, Malvern Panalytical Ltd, United Kingdom) at fixed angle of 173° at 25 °C. Samples were analyzed in triplicate. The same equipment was used for the determination of zeta potential.
7
Characterization of Silver Nanoparticles
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The bioreduction of silver ions was observed by visual assessment and measuring the UV-Vis spectra of the reaction medium within a wavelength of 250–1000 nm and was conducted using the LAMBDA 950 PerkinElmer UV/Vis/IR spectrophotometer (PerkinElmer Inc., Waltham, MA, USA). Zeta potentials of the prepared Ag-NPs were analyzed using Dynamic Light Scattering (DLS) (Zetasizer Nano ZN, Malvern Panalytical Ltd., (Malvern, WR14 1XZ, Malvern, UK) at a fixed angle of 173° at 25 °C. Using an EDX-equipped JSM-6510 (JEOL, Tokyo, Japan), energy dispersive X-ray (EDX) and a scanning electron microscope (SEM), morphological analyses were performed. To determine the crystalline structure and desired orientation of the prepared Ag-NPs, X-ray diffraction (XRD) measurements were captured by the EMPYREAN X-ray diffractometer with a wavelength (λ) of 0.15418 nm. The XRD patterns were documented at a scan speed of 2°/min by drop coating film on the glass substrate in a wide range of Bragg angles (30–80°).
8
Characterization of Selenium Nanoparticles Synthesized by P. chrysogenum
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The color change in the sodium selenite solution incubated with P. chrysogenum culture supernatant was clearly detected using UV-visible spectroscopy. The absorbance of the nanoparticle solution and the control sample was detected between 300 and 800 nm using a UV-visible spectrophotometer (PerkinElmer Life and Analytical Sciences, CT, Ohio, USA). To determine the size, shape, aggregation, and morphological characteristics of the SeNPs, transmission electron microscopy (TEM) images of high quality were collected using a TEM (HR-TEM; JEOL 2100, Japan). The FTIR spectra of the control medium and SeNP solutions were recorded using FTIR (PerkinElmer, Ohio, USA). All measurements were performed between 400 and 4000 cm−1. Dynamic light scattering (DLS) measurements using Zetasizer (Zetasizer Nano ZN, Malvern Panalytical Ltd., Malvern, UK) at an angle equal to 173° and a temperature of 25 °C were used to assess the particle size distribution and zeta potential of SeNPs. By measuring the polydispersity index (PDI), DLS analysis also revealed further details regarding the homogeneity of the NPs solutions. X-ray diffraction (XRD) analysis (Bruker D8 DISCOVER Diffractometer, USA) was used to assess the crystallinity of the myco-fabricated SeNPs.
9
Particle Size and Zeta Potential Analysis
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The particle size and size distribution in terms of the average volume diameters and polydispersity index of the prepared particles were analyzed via photon correlation spectroscopy using a particle size analyzer, dynamic light scattering (DLS) (Zetasizer Nano ZN, Malvern Panalytical Ltd, UK) at fixed angle of 173° at 25° C. Samples were analyzed in triplicate. The same equipment was used for the determination of zeta potential.
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