Live Cell imaging of Host-Virus interactions using CARS and Two-Photon microscopy
- Event time: 1:00pm
- Event date: 3rd May 2010
- Speaker: Christine Wong (Formerly School of Physics & Astronomy, University of Edinburgh)
- Location: Room 2511, James Clerk Maxwell Building (JCMB) James Clerk Maxwell Building Peter Guthrie Tait Road Edinburgh EH9 3FD GB
Confocal microscopy is an established imaging technique, however the addition of exogenous labels can disrupt cellular functions and often limits the technique to studying fixed samples. In contrast, coherent anti-Stokes Raman scattering (CARS) microscopy enables non-invasive intracellular imaging without the need for labelling. The CARS technique has been applied to a variety of biological specimens due to its properties of enhanced signal and chemical selectivity, enabling intracellular imaging with vibrational contrast at high acquisition speeds. In this way, CARS provides the capability for live cell imaging.
We have developed a multi-modal microscope platform that combines CARS and two-photon fluorescence microscopies in order to study the effects of viral infection on host cell morphology and lipid droplet distrubution. We apply this to study living fibroblast cells infected with cytomegalovirus. The virus is genetically modified to cause expression of the green fluorescent protein (GFP) in the host cell upon infection. The CARS laser source is tuned to the C?H stretch vibration at 2845 cm-1 in order to visualise the cell morphology and lipid droplets. The two-photon images reveal the extent of viral protein expression due to the amount of GFP expressed.
Therefore, using this microscope platform, we can show host-pathogen interactions by monitoring cellular changes, such as changes in lipid droplet distribution and nucleus morphology during live infection. This method has potential applications in the study of other host-virus interactions as well as the capability to study changes of cellular material other than lipid droplets.
This is a weekly series of informal talks given primarily by members of the soft condensed matter and statistical mechanics groups, but is also open to members of other groups and external visitors. The aim of the series is to promote discussion and learning of various topics at a level suitable to the broad background of the group. Everyone is welcome to attend..