293T cells were co-transfected with different SARS-CoV-2 spike plasmids with GFP1-10 plasmid (cat#68715, Addgene, USA) as effector cells. Another population of 293T cells was co-transfected with ACE2, TMPRSS2, and GFP11 (cat#68716, Addgene) as target cells. After 24 h post-transfection, the effector and target cells were digested by EDTA–Trypsin (25200-072, Gibco) and mixed at a 1:1 ratio. The mixed cells were co-cultured at a 37 °C incubator for another 24 h. The mixed cells were fixed in 10% formalin and then permeabilized with 0.1% Triton-X100 (Sigma, USA) at room temperature. The antifade mounting medium with 4′,6-Diamidino-2-Phenylindole, Dihydrochloride (DAPI, H-1200, Vector Laboratories) was used for mounting and DAPI staining. Images were taken with the Olympus BX73 fluorescence microscope (Olympus Life Science, Tokyo, Japan). The fusion area of images was quantified by ImageJ (v.1.8.0_345).
Bx73 fluorescence microscope
Manufactured by Olympus
Sourced in Japan
The BX73 is a fluorescence microscope designed for a variety of applications. It features an LED light source and a range of filter sets to enable fluorescence imaging. The microscope provides high-quality optical performance and is suitable for diverse research and analysis tasks.
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Lab products found in correlation
Gfp1 10 plasmid, by Addgene (2 mentions)
Gfp11, by Addgene (2 mentions)
Trypsin edta, by Thermo Fisher Scientific (2 mentions)
Triton x 100, by Merck Group (2 mentions)
Tmprss2, by Addgene (1 mentions)
Hace2, by Addgene (1 mentions)
4 6 diamidino 2 phenylindole dihydrochloride dapi h1200, by Vector Laboratories (1 mentions)
2 protocols using bx73 fluorescence microscope
1
SARS-CoV-2 Spike Protein Fusion Assay
2
SARS-CoV-2 Spike Protein Cell Fusion Assay
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293T cells were co-transfected with different SARS-CoV-2 spike plasmids with GFP1-10 plasmid (cat#68715, Addgene) as effector cells. Another population of 293T cells was co-transfected with human ACE2 (hACE2), TMPRSS2, and GFP11 (cat#68716, Addgene) as target cells. After 24 h post-transfection, the effector and target cells were digested by EDTA-Trypsin (25200072, Gibco) and mixed at a 1:1 ratio. The mixed cells were co-cultured at a 37°C incubator for another 24 h. The mixed cells were fixed in 10% formalin and then permeabilized with 0.1% Triton-X100 (11332481001, Sigma, USA) at room temperature. The antifade mounting medium with 4′,6-Diamidino-2-Phenylindole, Dihydrochloride (DAPI, H-1200, Vector Laboratories) was used for mounting and DAPI staining. Images were taken with the Olympus BX73 fluorescence microscope (Olympus Life Science, Tokyo, Japan). The fusion area of images was quantified by ImageJ.
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