Intracellular imaging of host-pathogen interactions using combined CARS and two-photon fluorescence microscopies.
Intracellular imaging is a key tool in the investigation of host-pathogen interactions. Advances in this area are particularly sought to understand the effect of viral infection processes on the host cell and its metabolic functions including those cases where host cell lipid metabolism is modulated as a result of infection. We demonstrate the use of combined coherent anti-Stokes Raman scattering (CARS) and two-photon fluorescence microscopies to image fibroblast cells infected by cytomegalovirus. CARS is used to image the host cell membrane, lipid droplets and morphology of the nucleus. Cell nuclei are found to expand during infection, approximately doubling in area. Some cells also show accumulations of lipid droplets at the nuclear periphery. Using a genetically modified virus strain expressing the green fluorescent protein also enables two-photon imaging of the same cells to reveal the location, nature and extent of viral protein expression.
Robinson
Iain
I
Collaborative Optical Spectroscopy, Micromanipulation and Imaging Centre COSMIC, School of Physics and Astronomy, The University of Edinburgh, Mayfield Road, Edinburgh EH9 3JZ, UK. iain@physics.org
Ochsenkühn
Michael Andreas
MA
Campbell
Colin J
CJ
Giraud
Gerard
G
Hossack
William J
WJ
Arlt
Jochen
J
Crain
Jason
J
D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D003586:Cytomegalovirus Infections / Q000175:diagnosis* / Q000378:metabolism / Q000821:virology; D003952:Diagnostic Imaging / Q000379:methods*; D015967:Gene Expression Regulation, Viral; D049452:Green Fluorescent Proteins; D054884:Host-Pathogen Interactions / Q000502:physiology*; D042541:Intracellular Space / Q000821:virology*; D050356:Lipid Metabolism; D008856:Microscopy, Fluorescence; D017785:Photons; D013059:Spectrum Analysis, Raman; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
D014764:Viral Proteins; D049452:Green Fluorescent Proteins