The presence of amyloid fibrils in the test samples was monitored by transmission electron microscopy (TEM). End-point solutions of peptides were collected and the samples were dissolved in the same solution to 0.2 mg/mL before studying with electron microscopy. A formvar-coated copper grid 400 mesh (Electron Microscopy Sciences, Hatfield, PA, USA) was placed on a 10 μL sample. After 5 min absorption, the grids with the preparation were negatively stained for 1.5–2.0 min with 1% (weight/volume) aqueous solution of uranyl acetate. The excess of the staining agent was removed with filter paper. The preparations were analyzed using a JEM-100C (Jeol, Tokyo, Japan) transmission electron microscope at the accelerating voltage of 80 kV. Images were recorded on the Kodak electron image film (SO-163) at nominal magnification of 40,000–60,000.
Electron image film so 163
Manufactured by Kodak
Sourced in United States, Japan
Electron image film (SO-163) is a high-resolution film designed for electron microscopy applications. It is a silver-halide based film with a specialized emulsion and backing that allows for the capture of high-quality images from electron microscopes. The film's primary function is to record and preserve electron microscope images for further analysis and documentation.
Automatically generated - may contain errors
Lab products found in correlation
Jem 100c, by JEOL (2 mentions)
Formvar coated copper grid 400 mesh, by Electron Microscopy Sciences (2 mentions)
Em uc7, by Leica (2 mentions)
Jem 2100 plus transmission electron microscope, by JEOL (2 mentions)
Epon 812, by Leica (2 mentions)
Epon 812, by SPI Supplies (2 mentions)
Tecnai f20, by Thermo Fisher Scientific (1 mentions)
6 protocols using electron image film so 163
1
Amyloid Fibrils Visualization by TEM
2
Negative Staining for TEM Visualization
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All the samples were dissolved in DMSO (the final concentration 5%) then the buffer (50 mM Tris–HCl, pH7.5) was added (0.2–0.5 mg/ml). Prior to negatively staining, the concentration of the samples was adjusted to 0.1–0.2 mg/ml. A copper grid (400 mesh) coated with a formvar film (0.2% ) was mounted on a sample drop (10μl). After 5–10 min absorption, the grid with the preparation was negatively stained for 1.5–2.0 min with 1% (weight/volume) aqueous solution of uranyl acetate. The excess staining agent was removed with filter paper. The preparations were analyzed using a JEM-1200 EX transmission electron microscope at the accelerating voltage of 80 kV. Images were recorded on the Kodak electron image film (SO-163) at nominal magnification of 40,000–60,000.
3
Characterizing Amyloid Peptide Morphology
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End-point solutions of peptides (0.2 mM in the buffer 50 mM Tris-HCl, pH 7.5, 150mM NaCl, 1% DMSO) incubated for 5 h with ThT were collected, and the samples were dissolved in the same solution to 0.2 mg/mL before studying with electron microscopy according to the previously described methods [55 (link),56 (link)], with minor modifications. A formvar-coated copper grid 400 mesh (Electron Microscopy Sciences, Hatfield, PA, USA) was placed on a 10 μL sample. After 5 min of absorption, the grid with the preparation was negatively stained for 1.5–2.0 min with a 1% (weight/volume) aqueous solution of uranyl acetate. The excess of the staining agent was removed with filter paper. The preparations were analyzed using a JEM-100C (Jeol, Tokyo, Japan) transmission electron microscope at the accelerating voltage of 80 kV. Images were recorded on the Kodak electron image film (SO-163) at a nominal magnification of 40,000–60,000.
4
Ultrastructural Analysis of Intestinal Samples
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Intestinal samples were fixed in glutaraldehyde and rinsed with 0.1 M phosphate buffer thrice for 15 min. Samples were then fixed in 1% osmium tetroxide for 2 h, dehydrated serially in 30% to 100% ethyl alcohol and dehydrated further with 100% acetone twice. The samples were then treated with mixture of acetone and Epon 812 (1:1; SPI Supplies, West Chester, PA, USA) for 4 h, incubated with acetone and Epon 812 (1:2) overnight, and embedded in pure Epon 812 for 5 h. Samples were then embedded in Epon 812 at 37 °C overnight, polymerized at 60 °C for 48 h, and sliced into 80-nm sections with an ultra-microtome (EM UC7, Leica, Wetzlar, Germany) before placing on cuprum grids. Cuprum grids were stained with uranium acetate and lead citrate and dried overnight at approximately 20 °C. Images were obtained using a JEM-2100Plus transmission electron microscope (JEOL, Tokyo, Japan) at 80 kV acceleration voltage and were recorded on Kodak Electron Image Film SO-163 (Eastman Kodak Co., Rochester, NY, USA).
5
Transmission Electron Microscopy Sample Preparation
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Muscle samples (0.1 cm × 0.1 cm × 0.1 cm) were fixed in glutaraldehyde and rinsed with 0.1 mol/L phosphate buffer thrice for 15 min. Samples were then fixed in 1% osmium tetroxide for 2 h, dehydrated serially in 30%–100% ethyl alcohol and dehydrated further with 100% acetone twice. The samples were then treated with acetone and Epon 812 (1:1; SPI Supplies, West Chester, PA, USA) for 4 h, incubated with acetone and Epon 812 (1:2) overnight, and embedded in pure Epon 812 for 5 h. Samples were then embedded in Epon 812 at 37 °C overnight, polymerized at 60 °C for 48 h, and sliced into 80 nm sections with an ultra-microtome (EM UC7, Leica, Wetzlar, Germany) before placing on cuprum grids. Cuprum grids were stained with uranium acetate and lead citrate and dried overnight at approximately 20 °C. Images were obtained using a JEM-2100Plus transmission electron microscope (JEOL, Tokyo, Japan) at 80 kV acceleration voltage and were recorded on Kodak Electron Image Film SO-163 (Eastman Kodak Co., Rochester, NY, USA).
6
Cryo-TEM Imaging Protocol
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Samples for cryo‐TEM investigations were prepared according to our standard method (see the Supporting Information). Microscopy was conducted using a FEI Tecnai F20 at a primary magnification of 100 000× (160 kV, FEG‐illumination) equipped with a FEI Eagle 2k CCD camera (pixel resolution 2.268 Å) or a Philips CM12 (100 kV, LaB6‐illumination) at a primary magnification of 58 300×. CM12 data were recorded on Kodak® electron image film SO 163 (Eastman Kodak Company, USA) and digitised for image processing to a final pixel resolution of 0.686 Å per pixel.
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