Negative stain electron microscopy was used to assess the different fibrillar deposits of patient-derived XFS samples. XFS materials were removed from the anterior lens capsule surface as described above in sample collection section. XFS patients were classified as having low, moderate, or heavy deposits. 6 μl of each sample was loaded onto a glow-discharged 400 mesh carbon-coated copper grid (Ted Pella Inc. Redding, CA). Typically, samples were adsorbed onto the grid surface for 2 min after which excess solution was removed by blotting with Whatman paper. Grids were then washed briefly and negatively stained using freshly filtered 2% uranyl acetate (UA) or 2% phosphotungstic acid (PTA) solutions. The negatively stained samples were analyzed with a FEI Tecnai G20 electron microscope (FEI Company). An acceleration voltage of 200 kV was used to record micrographs on an Eagle 4 k × 4 k CCD camera (FEI Company).
Fei tecnai g20 electron microscope
Manufactured by Thermo Fisher Scientific
The FEI Tecnai G20 is a transmission electron microscope (TEM) designed for high-resolution imaging and analysis of materials at the nanoscale. It is capable of producing detailed, magnified images of samples by transmitting a beam of electrons through a thin specimen. The core function of the FEI Tecnai G20 is to enable researchers and scientists to visualize and study the microstructure and composition of a wide range of materials with high resolution and accuracy.
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2 protocols using fei tecnai g20 electron microscope
1
Negative Stain Electron Microscopy of XFS
2
Negative Staining of Amyloid-Beta Samples
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The Aβ samples with or without Mimo-AuNPs (10 μL) were loaded onto freshly glow-discharged 200 mesh carbon-coated copper grids and adsorbed onto grid surface for ~1min. Grids were then washed with 50 μL 0.1 M and 0.01 M ammonium acetate and negatively stained with 2% phosphotungstic acid solution. After drying, the stained samples were analyzed with a FEI Tecnai G20 electron microscope (FEI Company, The Netherlands). An acceleration voltage of 200 kV was used to record micrographs on an Eagle 4k x 4k CCD camera.
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