Author: Van der Gucht, Winke; Stobbelaar, Kim; Govaerts, Matthias; Mangodt, Thomas; Barbezange, Cyril; Leemans, Annelies; De Winter, Benedicte; Van Gucht, Steven; Caljon, Guy; Maes, Louis; De Dooy, Jozef; Jorens, Philippe; Smet, Annemieke; Cos, Paul; Verhulst, Stijn; Delputte, Peter L.
Title: Isolation and Characterization of Clinical RSV Isolates in Belgium during the Winters of 2016–2018 Document date: 2019_11_6
ID: 0imlae98_19
Snippet: One day prior to inoculation, HEp-2 cells were seeded at a concentration of 17,500 cells/well in black CELLSTAR ®® 96-well plates with a µclear ®® flat bottom suitable for fluorescence microscopy (Greiner bio-one). Cells were inoculated with clinical RSV and RSV A2 at a MOI of 0.05 for 2 h at 37 • C (5% CO 2 ). After 2h, the inoculum was removed and replaced by DMEM-10 containing 0.6% Avicel ®® (FMC biopolymer). After 48 h cells were was.....
Document: One day prior to inoculation, HEp-2 cells were seeded at a concentration of 17,500 cells/well in black CELLSTAR ®® 96-well plates with a µclear ®® flat bottom suitable for fluorescence microscopy (Greiner bio-one). Cells were inoculated with clinical RSV and RSV A2 at a MOI of 0.05 for 2 h at 37 • C (5% CO 2 ). After 2h, the inoculum was removed and replaced by DMEM-10 containing 0.6% Avicel ®® (FMC biopolymer). After 48 h cells were washed with PBS, fixed with 4% paraformaldehyde solution, permeabilized with triton X-100 and stained with polyclonal goat-anti-RSV (Virostat; Westbrook, ME, USA) followed by donkey-anti-goat secondary antibody conjugated with AF488 (Thermo Fisher Scientific). DAPI staining was performed to stain the nuclei (Sigma-Aldrich).
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