To determine the effect of P9R on viral particles, SARS-CoV-2 was pretreated by 100 μg ml−1 of P9R or P9SHN for 1 h. The virus was fixed by formalin for overnight and then applied to continuous carbon grids. The grids were transferred into 4% uranyl acetate and incubated for 1 min. After removing the solution, the grids were air-dried at room temperature. For each peptide/DNA nanoparticle, three independent experiments were done for taking TEM images by FEI Tecnal G2-20 TEM.
G2 20 tem
Manufactured by Thermo Fisher Scientific
The G2-20 TEM is a transmission electron microscope (TEM) manufactured by Thermo Fisher Scientific. It is designed to provide high-resolution imaging and analysis of materials at the nanoscale level. The core function of the G2-20 TEM is to generate and transmit a beam of electrons through a thin specimen, creating an image that can be magnified and observed on a fluorescent screen or captured digitally.
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3 protocols using g2 20 tem
1
Evaluating SARS-CoV-2 Viral Particles
2
Analyzing FBP's Effect on Viral Particles
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To determine the effect of FBP on viral particles, the A(H1N1) virus was treated with 200 μg ml−1 of FBP, PBS, or Triton X-100 (0.15%) for 1 h. The virus was fixed by formalin overnight and then applied to continuous carbon grids. The grids were transferred into 4% uranyl acetate and incubated for 1 min. After removing the solution, the grids were air-dried at room temperature. For each sample, two to three biological samples were done for taking TEM images by FEI Tecnal G2-20 TEM.
3
Evaluating P16 Inhibition of Viral Particles
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To determine the effect of P16 on viral particles, A(H1N1) virus was pretreated by 200 μg ml -1 of P16, PBS or Triton X-100 (0.15%) for 1h. The virus was xed by formalin for overnight and then applied to continuous carbon grids. The grids were transferred into 4% uranyl acetate and incubated for 1 min. After removing the solution, the grids were air-dried at room temperature. For each sample, two-three biological samples were done for taking TEM images by FEI Tecnal G2-20 TEM.
H5N1 pseudovirus assay H5N1 pseudotype virus(38) bearing H5N1 HA and NA was pretreated with PBS or P16 in PBS and then incubated at RT for 1h. MDCK cells were infected with the treated pseudotype virus for 1h. MDCK cells without pseudotype virus infection were served as the baseline control of luciferase protein. After 18h cell culture, cell lysates were collected and the luciferase protein was measured by Luciferase assay system (Promega) in a Victor X3 Multilabel reader (PerkinElmer). The luminescence reading was normalized to 1mg protein.
H5N1 pseudovirus assay H5N1 pseudotype virus(38) bearing H5N1 HA and NA was pretreated with PBS or P16 in PBS and then incubated at RT for 1h. MDCK cells were infected with the treated pseudotype virus for 1h. MDCK cells without pseudotype virus infection were served as the baseline control of luciferase protein. After 18h cell culture, cell lysates were collected and the luciferase protein was measured by Luciferase assay system (Promega) in a Victor X3 Multilabel reader (PerkinElmer). The luminescence reading was normalized to 1mg protein.
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