Technical Reference #1899

1899.

Genetic engineered, biarsenical labelled influenza virus allows visualization of viral NS1 protein in living cells Yang Li, Xinya Lu, Junwei Li, Nathalie Bérubé, Kerri-Lane Giest, Qiang Liu, Deborah H. Anderson, and Yan Zhou, University of Saskatchewan, Journal of Virology, (2010)
Link To Paper

Materials & Methods:
For visualizing NS1 nuclear import, A549 cells seeded in glass-bottom 35 mm dishes (MatTek) were infected with PR8-411 at an MOI of 1. At 3 h.p.i., the media were changed to Opti-MEM containing FlAsH (0.2 μM), EDT (50 μM) and β- mercaptoethanol (10 μM). Between 4 and 7 h.p.i, live-cell fluorescence images of infected A549 cells were recorded every 3 min using a Leica SP5 microscope equipped with a 40 × NA 1.25 oil objective, an I3 filter cube, a PB450-490 excitation filter and an LP515 emission filter. All images were acquired with 500-ms exposures under the same illumination conditions. Images were analyzed using Leica LAS AF 1.8.2 software

Microscopic Technique
Live-cell Fluorescence

Cell Type(s)
A549 cells