Journal
ACS NANO
Volume 11, Issue 3, Pages 2575-2585Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acsnano.6b05601
Keywords
coherent brightfield microscopy; high-speed imaging; single-virus tracking; early stage infection; virus-membrane interaction; diffusion; 3D localization; interferometric imaging; digital background removal; local delivery of virus particle
Categories
Funding
- Nano Program of Academia Sinica
- Ministry of Science and Technology, Taiwan [102-2112-M-001-002-MY3]
- Academia Sinica Postdoctoral Research Fellowship
Ask authors/readers for more resources
Viral infection starts with a virus particle landing on a cell surface followed by penetration of the plasma membrane. Due to the difficulty of measuring the rapid motion of small-sized virus particles on the membrane, little is known about how a virus particle reaches an endocytic site after landing at a random location. Here, we use coherent brightfield (COBRI) microscopy to investigate early stage viral infection with ultrahigh spatiotemporal resolution. By detecting intrinsic scattered light via imaging-based interferometry, COBRI microscopy allows us to track the motion of a single vaccinia virus particle with nanometer spatial precision (<3 nm) in 3D and microsecond temporal resolution (up to 100,000 frames per second). We explore the possibility of differentiating the virus signal from cell background based on their distinct spatial and temporal behaviors via digital image processing. Through image postprocessing, relatively stationary background scattering of cellular structures is effectively removed, generating a background-free image of the diffusive virus particle for precise localization. Using our method, we unveil single virus particles exploring cell plasma membranes after attachment. We found that immediately after attaching to the membrane (within a second), the virus particle is locally confined within hundreds of nanometers where the virus particle diffuses laterally with a very high diffusion coefficient (similar to 1 mu m(2)/s) at microsecond time scales. Ultrahigh-speed scattering-based optical imaging may provide opportunities for resolving rapid virus receptor interactions with nanometer clarity.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available