Author: Meier, Anita F.; Suter, Mark; Schraner, Elisabeth M.; Humbel, Bruno M.; Tobler, Kurt; Ackermann, Mathias; Laimbacher, Andrea S.
Title: Transfer of Anti-Rotavirus Antibodies during Pregnancy and in Milk Following Maternal Vaccination with a Herpes Simplex Virus Type-1 Amplicon Vector Document date: 2017_2_16
ID: 09hmet7r_62
Snippet: HepG2 cells were transduced (MOI 5) with the HSV-1 amplicon vector sWa[VP2/6/7_V5] and 24 hpt, the cells were scraped into the medium and centrifuged for 20 min at 4000× g. The resulting cell pellet was fixed with 4% formaldehyde in 0.1 M Na/K phosphate buffer for 4 h. Thereafter, the pellet was dehydrated with ascending ethanol series starting at 70%, followed by 80%, 96% and three times in absolute ethanol for 10 min each. Next, the pellet was.....
Document: HepG2 cells were transduced (MOI 5) with the HSV-1 amplicon vector sWa[VP2/6/7_V5] and 24 hpt, the cells were scraped into the medium and centrifuged for 20 min at 4000× g. The resulting cell pellet was fixed with 4% formaldehyde in 0.1 M Na/K phosphate buffer for 4 h. Thereafter, the pellet was dehydrated with ascending ethanol series starting at 70%, followed by 80%, 96% and three times in absolute ethanol for 10 min each. Next, the pellet was incubated at 4 • C for 1.5 h in a 2:1 mix of LR White/Ethanol followed by infiltration of LR White (EMS) alone for 4 h and a final change of LR White followed by overnight incubation at 4 • C. Embedding in LR white was done in gelatin capsules at 50 • C for 24 h in an oven. Ultrathin sections were cut and collected on carbon-coated Formvar films mounted on single slot copper grids (2 × 1 mm; EMS).
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