Journal: bioRxiv

Article Title: Enhanced Visualization of Influenza A Virus Entry Using Virus-View Atomic Force Microscopy

doi: 10.1101/2024.07.19.603848

Figure Lengend Snippet: Live MDCK cells expressing EGFP-Lifeact were imaged by ViViD-AFM at 27°C at 10 s intervals. (A) Representative morphology image of the cell membrane (left), fluorescence image of Lifeact (middle), and merged image (right). Scale bar: 1000 nm. (B) Cross-sectional profile of the membrane morphology (gray) and Lifeact fluorescence intensity (green) along the dashed line in panel (A). The lowest height was set as 0 nm. (C) Pearson’s correlation coefficient (r) between height of membrane morphology (x axis) and Lifeact fluorescence intensity (y axis), for the profile data in panel (B). AU, arbitrary units.

Article Snippet: For transient expression of Lifeact-EGFP, the pFX-Lifeact-EGFP expression vector was transfected into MDCK cells using Polyethylenimine Max (Polysciences, Warrington, PA, USA).

Techniques: Expressing, Membrane, Fluorescence

Journal: bioRxiv

Article Title: Enhanced Visualization of Influenza A Virus Entry Using Virus-View Atomic Force Microscopy

doi: 10.1101/2024.07.19.603848

Figure Lengend Snippet: MDCK cells transiently expressing EGFP-Lifeact were inoculated with IAV-AF594 at 27°C and subjected to 10 s interval time-lapse ViViD-AFM imaging. (A) Live imaging of IAV internalization by ViViD-AFM. From top to bottom, sequential images of AFM, IAV-AF594 (magenta), EGFP-Lifeact (green), the merge of AFM and IAV-AF594, and the merge of AFM and EGFP-Lifeact are displayed every 30 s. t = 0 s was set as the moment when the virion morphology was no longer detected. Image size: 1500 × 1500 nm 2 . (B) Correlation analysis of membrane morphology and fluorescence, related to panel (A). From top, sequential images of AFM, the merge of AFM and IAV-AF594 (magenta), and the merge of AFM and the EGFP-Lifeact (green) at t = −10 s, 0 s, and 10 s are displayed. The lowest panel is a plot of the cross-sectional profile of membrane morphology and Lifeact fluorescence intensity along the dashed line in the above merged image. (A, B) Arrowheads: virion morphology (yellow), membrane ruffles (blue) and virion fluorescence (orange). (C) Pearson’s correlation coefficient (r) between height of membrane morphology (x axis) and Lifeact fluorescence intensity (y axis), for the profile data in panel (B).

Article Snippet: For transient expression of Lifeact-EGFP, the pFX-Lifeact-EGFP expression vector was transfected into MDCK cells using Polyethylenimine Max (Polysciences, Warrington, PA, USA).

Techniques: Expressing, Imaging, Membrane, Fluorescence