Regensburg 2013 – scientific programme

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BP: Fachverband Biologische Physik

BP 10: Posters: Imaging

BP 10.12: Poster

Monday, March 11, 2013, 17:30–19:30, Poster B2

Stimulated Raman Scattering for noninvasive live imaging — •Michael Stührenberg, Henning Hachmeister, Lena Nolte, and Thomas Huser — Biomolecular Photonics, Universität Bielefeld, Germany

A common method for the visualisation of biological molecules is fluorescence microscopy. By using a fluorescent dye, the molecules of interest can be made visible. Fluorescence microscopy covers a wide range of techniques and continues to gain popularity in respect to i.e. superresolution and 3D live cell imaging. Nevertheless, these techniques have some major disadvantages. Amongst other things there is no guarantee that the probes won't affect certain characteristics of the molecule, such as motility or binding capability. Therefor, the use of fluorescent probes requires careful controls in order for it to not be detrimental to biological processes. To overcome these drawbacks, we demonstrate a rapid noninvasive imaging technique based on Raman scattering. In Stimulated Raman Scattering (SRS) the molecule of interest is excited to a specific vibrational mode. This is achieved by the application of 2 different confocal laserbeams overlapping in time and space. Through excitation in a vibrational mode, one beam loses intensity, while the other one experiences, through induced stimulation, an intensity gain. The probability for this is small compared to a fluorescent signal and requires a lock-in based detection system. Yet this results in a nearly background free noninvasive imaging technique, which can be tuned to a wide range of molecules. The ultimate goal is live imaging for which we apply a homebuilt lock-in amplifier with a high temporal resolution.