Ultrathin carbon film on lacey carbon support with 400 mesh on copper (Ted Pella, 01824) was glow discharged for 30 s using PELCO easiGlow glow discharger (Ted Pella). Isolated particle-enriched fractions from healthy individuals were put on charged grids for 1 min, washed for 30 s with MilliQ water, and fixed with 1% uranyl acetate for 30 s. Grids with stained samples were air dried for at least 30 min before imaging. Prepared samples were imaged at 120 kV using FEI Tecnai™ Spirit TEM system. FEI- Tecnai™ Spirit TEM system was interfaced to a bottom mounted Eagle™ 2 K TEM CCD multiscan camera and to a NanoSprint12S-B cMOS camera from Advanced Microscopy Techniques (AMT) fast side mounted TEM CCD Camera. Images were collected at 30,000x magnification under 3 μm defocus. Images were acquired as 4000 × 3000 pixel, 16-bit gray scale files using the Nanosprint 12 AMT interface and camera. For the analysis, TEM images were acquired with at least 25 particles were measured from each plasma fraction. Observed particles per fraction were binned with 5 nm width for the histogram. Particle diameters were manually measured using Fiji line-tool, and exported to excel/R for downstream analyses.
Fei tecnai spirit tem system
Manufactured by Thermo Fisher Scientific
The FEI Tecnai™ Spirit TEM system is a transmission electron microscope designed for high-resolution imaging and analysis of samples at the nanoscale level. It provides a comprehensive solution for advanced materials characterization and biological research.
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2 protocols using fei tecnai spirit tem system
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Transmission Electron Microscopy Imaging of Extracellular Vesicles
2
Platelet Imaging via Transmission Electron Microscopy
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Ultrathin carbon lm on lacey carbon support with 400 mesh on copper (Ted Pella, catalog number: 01824) was glow discharged for 30 seconds using PELCO easiGlow glow discharger (Ted Pella). Isolated platelet samples (RT vs. -80°C) were put on charged grids for 1 min, washed for 30 seconds with MilliQ water, and xed with 1% uranyl acetate for 30 seconds. Grids with stained samples were air dried at least 30 minutes before imaging. Prepared samples were imaged at 120 kV using FEI Tecnai™ Spirit TEM system. FEI-Tecnai™ Spirit TEM system was interfaced to a bottom mounted Eagle™ 2K TEM CCD multiscan camera and to a NanoSprint12S-B cMOS camera from Advanced Microscopy Techniques (AMT) fast side mounted TEM CCD Camera. Images were collected at 8,000-80,000x magni cation under 1-2μm defocus. Regions shown in Figure 3 were representative of each sample, which were uniformly observed from larger elds of view across the grids. Images were acquired as 2048 × 2048 pixel, 16-bit gray scale les using the FEI's TEM Imaging & Analysis (TIA) interface on an Eagle™ 2K CCD multiscan camera.
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