A Varian Cary Eclipse (Cary 50) UV-vis spectrophotometer was used to carry out the optical properties of the NPs. The photoluminescence spectra of the NPs were recorded on a Varian Cary Eclipse EL04103870 spectrofluorometer with a medium PMT voltage at an excitation wavelength of 200 nm. The absorption and emission spectra were obtained in chloroform and placed in quartz cuvettes (1 cm path length). Powdered XRD patterns of the samples were measured on a Bruker MeasSrv D2-205530 diffractometer using secondary graphite monochromated Cu Kα radiation (λ 1.5406 Å) at 30 kV/30 mA. Measurements were taken using a glancing angle of incidence detector at an angle of 2°, for 2θ values over 10–90° in steps of 0.026° with a step time of 37 s and at a temperature of 25 °C. X-ray photoelectron spectroscopy measurements were performed with a PHI 5000 Versaprobe – Scanning ESCA Microprobe operating with a 100 μm 25 W 15 kV Al monochromatic X-ray beam. The transmission electron microscopy (TEM) was carried out on a FEI Technai T12 operated at an acceleration voltage of 200 kV with a beam spot size of 20–100 nm in TEM mode. The vibrational modes were measured on a Bruker Tensor 27 FT-IR and the nuclear magnetic resonance (NMR) data was obtained using a 500 MHz Bruker AVANCE III, the samples were run using CDCl3 as a solvent at 300 K.
Technai t12
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Technai T12 is a transmission electron microscope (TEM) designed for high-resolution imaging and analysis of samples. It is equipped with a LaB6 electron source, providing a stable beam of electrons for advanced imaging and analytical capabilities. The Technai T12 is capable of achieving a resolution of up to 0.2 nanometers, enabling the visualization of fine structural details in a wide range of materials.
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4 protocols using technai t12
1
Comprehensive Characterization of Nanoparticles
2
Microwave-Assisted TEM Wing Preparation
3
Synthesis of Citrate-Stabilized Gold Nanoparticles
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Example 2
All glassware used for AuNPs synthesis are cleaned in NOCHROMIX solution (Godax Laboratories, Inc., Cabin John, Md.) and then aqua regia (3 parts HCL+1 part HNO3) according to standard lab procedure. The synthesis of citrate-stabilized gold nanoparticle (AuNP) was based on a modification of Turkevich method. (Enustum et al., 1963, J. Am. Chem. Soc. 85(21):3317-3328). Briefly, 100 mL solution of 1 mM HAuCl4 (Sigma-Aldrich, St. Louis, Mo.) was boiled with stirring until bubbles formation and having a uniform temperature. Heating was kept for a further 25 minutes. Then, 10 mL of preheated trisodium citrate (38.8 mM, Sigma-Aldrich, St. Louis, Mo.) was added to the boiling HAuCl4 solution quickly. The solution turned to colorless for a moment, followed by violet to dark ruby red. The solution was heated for a further five minutes before cooling to room temperature. The solution was stored in the dark in a flask covered with foil. The sizes of AuNPs were characterized to be 32±3 nm using transmission electron microscopy (TECHNAI T12, FEI, Hillsboro, Oreg.).
4
Visualization of MAX1 Peptide Fibrils
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1 wt.% MAX1 gel or 1 wt.% MAX1 gel with 10 mM dopamine were prepared in Eppendorf tubes as described above and incubated at 37 °C for three days. To allow visualization of distinct fibers, gel samples were diluted 50× into water. A 5 µL drop of the diluted peptide solution was placed on a 200 mesh copper grid covered by carbon film (Electron Microscopy Science, Hatfield, PA, USA) for 1 min and blotted by filter paper. Subsequently, for washing, 5 µL of water were added to the grid for several s and blotted by filter paper. Immediately after that, 0.75% uranyl formate was added to the grid and allowed to stand for 1 min, then blotted with a filter paper and left to air dry. Images were taken with a Technai T12 (FEI Company, Hillsboro, OR, USA) at 80 kv accelerating voltage. Average fibril width was measured via ImageJ software (National Institutes of Health, Bethesda, MD, USA) by taking 80 (MAX1) or 193 (MAX1/dopamine) independent measurements from distinct fibrils in the field of view of fibrils observed at 3 separate micrographs, representing different location of the fibrils.
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