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AKB: Biologische Physik

AKB 30: Poster Session I

AKB 30.38: Poster

Montag, 27. März 2006, 15:30–18:00, P1

Dual-Color Single-Virus Tracing: Investigating the Entry Pathway of HIV — •Stefan Riegelsberger¹, John A. G. Briggs¹, Barbara Müller², Marko Lampe², Thomas Endress¹, Don C. Lamb¹, Hans-Georg Kräusslich², and Christoph Bräuchle¹ — ¹Physical Chemistry, Department of Chemistry and Biochemistry, Ludwig-Maximillians-Universität München, Butenandtstr. 11, Haus E, 81377 München, Germany — ²Abteilung Virologie, Universitätsklinikum Heidelberg, Im Neuenheimer Feld 324, 69120 Heidelberg, Germany

Detailed information about the interactions between virus particles and living cells can be obtained by visualising and tracking individual virus particles in real time. For the investigation of HIV entry, we have developed a novel, dual-colour HIV derivative that allows us to distinguish between complete viruses and those that have undergone fusion. We are observing the infection pathway of these particles using a dual-colour microscope. The microscope is based on wide-field microscopy with laser excitation and ultrasensitive fluorescence detection. The laser beams are guided through a vibrating, multimode optical fiber to remove interference fringes in the image plane, creating a very homogenous excitation profile. Improved depth resolution is achieved using a Köhler illumination scheme. The excitation sources are interleaved such that green and red fluorophores are excited on alternate images, and the emitted light is further spectrally separated onto two regions of the camera. This set up removes cross-talk between the two fluorescent labels, and allows us to distinguish broad-spectrum cellular autofluorescence from the viral signals and to correct for it.