Nanoparticles zeta potential and hydrodynamic diameter were measured using 0.01 mg mL−1 water suspensions in DST1070 folded cuvette in a Zetasizer Nano-ZS (Malvern). Size and shape were determined by Transmission Electron Microscopy in a Carl Zeiss CEM-902 with 80 keV using copper grids and parlodium film. The microscope was equipped with a Castaing-Henry-Ottensmeyer filter spectrometer used for the electron energy loss spectrometry (EELS). The crystal structure was determined by X-ray Diffraction (XRD) using a Shimadzu XRD7000 operating with Cu-Kα (λ = 0.154060 nm), in continuous mode with a 2° min−1 speed from 20° to 80°. Surface functionalisation was investigated by Infrared Spectroscopy equipped with Attenuated Total Reflectance (ATR-FTIR) using an Agilent Cary 630. X-Ray Photoelectron spectra were obtained using a SPECS system (SPECS GmbH) equipped with X-ray XR-50 with radiation Al Kα (hν = 1486.6 eV) and Phoibos 100 analyser with MCD-9 detector at the CCS Nano laboratory (Centro de Componentes Semicondutores, UNICAMP). Thermogravimetric Analysis (TGA) was performed using a STD q600 (TA Instruments), with a 10° min−1 speed in a nitrogen atmosphere.
Cem 902
Manufactured by Zeiss
The CEM 902 is a transmission electron microscope (TEM) manufactured by Zeiss. It is designed to provide high-resolution imaging and analytical capabilities for a variety of materials and samples. The CEM 902 is capable of performing electron diffraction, electron energy loss spectroscopy (EELS), and other advanced techniques.
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Lab products found in correlation
10 protocols using cem 902
1
Comprehensive Nanoparticle Characterization
2
Characterization of Engineered Vesicles
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Samples in PBS buffer were applied to 400-mesh carbon-coated copper grids (Electron Microscopy Sciences), stained with 2% uranyl acetate, air dried, and visualized on a Zeiss CEM 902 transmission electron microscope operating at voltage of 120 kV. Dynamic light scattering (DLS) for characterization of size of engineered vesicles was performed on a ZetaPALS (Brookhaven Instruments, Holtsville, NY). Collected vesicles were suspended in PBS buffer, and 1 ml of the vesicle suspension was transferred to a square cuvette for DLS measurement. DLS evaluates the time-dependent fluctuations of spherical particles in a light scattering intensity caused by the Brownian motion and a hydrodynamic diameter is calculated via the Stokes-Einstein equation [22] (link). The diameter of vesicles was calculated using BTC particle sizing software (Brookhaven Instruments, Holtsville, NY), demonstrating an average particle size of about 50 nm.
3
TEM Characterization of Nanomaterial Assemblies
4
TEM Preparation of Cell Fractions
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HCT-15 cells were fractionated as described above. Cellular and subcellular fractions were fixed in 3% glutaraldehyde (PBS, pH 7.3) for two days, washed with sodium phosphate buffer (pH 7.3), post-fixed with 1% osmium tetroxide for 1 h, and dehydrated through a series of graded alcohol baths. Standard TEM preparation techniques were used to obtain sample blocks that were cut in ultra-thin sections (80 nm) for mounting onto 100 mesh copper grids. The grids were stained with 2% alcoholic uranyl acetate and Reynold’s lead citrate. The grids were examined with a Zeiss CEM 902 transmission electron microscope, and images were captured using an AMT602 digital camera.
5
Insulin Localization in Pancreatic Islets
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Immunoelectron microscopy was performed as described previously (35 (link)). Briefly, islets were cultured overnight in the absence of glucose and then stimulated with 14 mmol/L glucose for 5 min and fixed in 4% paraformaldehyde and 0.5% glutaraldehyde in 0.1 mol/L phosphate buffer at room temperature. The fixed islets were dehydrated in graded ethyl alcohol and placed in a mixture of acrylic resin (LR White: 70% alcohol, 1:2) for 1 h and then in pure acrylic resin for 48 h at room temperature. Ultrathin 70-nm sections were mounted on 200-mesh nickel grids and stained using protein A–gold technique (36 (link)–38 (link)). Sections were stained with a guinea pig anti-mouse insulin antibody (1:5,000) diluted in PBS, 1% bovine albumin, and 1% glycine, at pH 7.4, followed by washing and immersion for 1 h in a solution of 15-nm diameter colloidal gold particles (CGP) covered with protein A (Janssen Pharmaceutica, Olen, Belgium) at a dilution of 1:50 (pH 8.2) in a moist chamber at 37°C. The grids were subjected to immunoelectron microscopy (Zeiss CEM 902).
6
TEM Analysis of Nanoparticle Assemblies
7
Asiaticoside Effects on Ultrastructure
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For electron microscopy, the 0, 6, 12, and 24 μM Asiaticoside-treated cells were fixed in a solution of 4% glutaraldehyde 0.05 M sodium cacodylate, postfixed in 1.5% OsO4, and dehydrated in alcohol (70%). They were then prepared for flat embedding in Epon 812 and then observed using a Zeiss CEM 902 electron microscope.
8
Autophagy Evaluation in SCC Cancer Cells
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Autophagy in transfected SCC-1 and SCC-9 cancer cells was investigated via transmission electron microscopy (TEM) analysis. Cells were harvested at 80% growth confluence, and then subjected to trypsinization. Afterwards, cells were first fixed by glutaraldehyde (2%) in phosphate buffer (0.1 M) followed by post fixation with osmium tetroxide (1%). Cells were then exposed to ethanol embedded in resin followed by cutting of thin sections with an ultramicrotome. Finally, these sections were examined on the transmission electron microscope (Zeiss CEM 902) linked to a digital camera for capturing different fields.
9
Electron Microscopy of Mouse Kidneys
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For electron microscopy of mouse kidneys, the samples were fixed in a solution of 2% glutaraldehyde 0.1 M sodium cacodylate, post-fixed in 1% OsO4, and dehydrated in alcohol. They were then processed for flat embedding in Epon 812 and observed in a Zeiss CEM 902 electron microscope.
10
Electron Microscopy Sample Preparation
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For electron microscopy, the cells were fixed in a solution of 4% glutaraldehyde 0.05 M sodium cacodylate, postfixed in 1.5% OsO4, and dehydrated in alcohol. They were then prepared for flat embedding in Epon 812 and then observed using a Zeiss CEM 902 electron microscope.
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