Approximately 5 μl volumes of each formaldehyde-fixed poliovirus sample were placed on carbon-stabilized, formvar-coated 400-mesh copper grids (Electron Microscopy Sciences, Hatfield, PA) for 5 min. Thereafter, the grids were gently washed with water followed by addition of 5 μl aqueous 2% uranyl acetate for 1 min. The grids were then wicked off, air-dried and examined in a JEOL JEM-1011 transmission electron microscope (JEOL USA, Inc., Peabody, MA) and images were recorded on an AMT XR50 digital camera (Advanced Microscopy Techniques, Corp. of Woburn, MA). Virion particles that closely resembled unirradiated virus were counted by visual (manual) enumeration of virion images on multiple micrographs. Virion particles that more closely resembled broken or empty particles were also counted by visual enumeration of virus images. In each sample >700 particles were analyzed and counted.
Formvar coated 400 mesh copper grids
Manufactured by Electron Microscopy Sciences
Sourced in United States
Formvar-coated 400-mesh copper grids are a type of specimen support used in electron microscopy. They consist of a copper grid with a thin film of the polymer Formvar coated onto the surface, providing a support structure for samples to be examined under an electron microscope.
Automatically generated - may contain errors
Lab products found in correlation
Polymethyl methacrylate, by McMaster-Carr (2 mentions)
Nucleopore hydrophilic membrane filters, by Cytiva (2 mentions)
Thermo fisher total exosomal rna, by Thermo Fisher Scientific (2 mentions)
Thermo fisher taqman mirna probes, by Thermo Fisher Scientific (2 mentions)
Aqueous 2 uranyl acetate, by Electron Microscopy Sciences (2 mentions)
Protein extraction kit, by Thermo Fisher Scientific (2 mentions)
Whatman, by Cytiva (2 mentions)
Jem 1011, by JEOL (1 mentions)
Cm12 transmission electron microscope, by Philips (1 mentions)
6 protocols using formvar coated 400 mesh copper grids
1
Transmission Electron Microscopy of Poliovirus
2
Visualizing Viral Matrix Protein Interactions
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MeV-M and NiV-M for single-particle analysis were diluted to a concentration of 0.02 mg/ml before negative staining. For incubation with lipid vesicles, purified MeV-M or NiV-M (10 μM) was incubated with the indicated lipids (400 μM) for 3 days at 37°C. Three microliters of sample was each applied to freshly plasma-cleaned carbon and formvar-coated 400-mesh copper grids (Electron Microscopy Sciences). After 1 min, excess liquid was wicked off and the grids were briefly washed on droplets of double-distilled water, followed by two brief washes on droplets of a 2% (v/v) aqueous uranyl acetate solution and lastly incubated on a 2% (v/v) aqueous uranyl acetate solution for 1 min. Excess stain was removed and the grids were dried thoroughly. Each sample was examined on an FEI Titan Halo 300-kV electron microscope with a Falcon 3EC camera. M proteins were imaged at ×75,000 magnification, and M proteins with liposomes were imaged at ×96,000 magnification.
3
Negative Staining of Virus Particles
4
Exosome Isolation and Characterization
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Polymethyl methacrylate (PMMA) with a thickness of 1.5 mm and Double-Sided Adhesive (DSA) were purchased from McMaster–Carr (Atlanta, GA, USA). Whatman Nucleopore Hydrophilic Membrane filters with 2.0 μm pore size, 30 nm pore size, and 15 nm pore size, and with a 25 mm diameter circle each, 1X Phosphate-buffered Saline (PBS), Thermo Fisher’s Total Exosome Isolation Kit (from plasma), Thermo Fisher Total Exosomal RNA, and Protein Extraction Kit were all obtained from Fischer Scientific (Fair lawn, NJ, USA). The Thermo Fisher Taqman miRNA probes were purchased from Thermo Fisher Scientific (Waltham, MA, USA). The 400 mesh Formvar-coated copper grids and the aqueous 2% uranyl acetate were obtained from Electron Microscopy Sciences (Hatfield, PA, USA).
5
Exosome Isolation and Characterization
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Polymethyl methacrylate (PMMA) with a thickness of 1.5 mm and Double-Sided Adhesive (DSA) were purchased from McMaster–Carr (Atlanta, GA, USA). Whatman Nucleopore Hydrophilic Membrane filters with 2.0 μm pore size, 30 nm pore size, and 15 nm pore size, and with a 25 mm diameter circle each, 1X Phosphate-buffered Saline (PBS), Thermo Fisher’s Total Exosome Isolation Kit (from plasma), Thermo Fisher Total Exosomal RNA, and Protein Extraction Kit were all obtained from Fischer Scientific (Fair lawn, NJ, USA). The Thermo Fisher Taqman miRNA probes were purchased from Thermo Fisher Scientific (Waltham, MA, USA). The 400 mesh Formvar-coated copper grids and the aqueous 2% uranyl acetate were obtained from Electron Microscopy Sciences (Hatfield, PA, USA).
6
Electron Microscopy of α-Synuclein Fibrils
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