Regensburg 2013 – wissenschaftliches Programm

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BP: Fachverband Biologische Physik

BP 28: Focus session: Intracellular Spectroscopy

BP 28.4: Vortrag

Donnerstag, 14. März 2013, 10:30–10:45, H44

Labelfree coherent Raman scattering microspectroscopy: A new intracellular spectroscopy tool in lipidomic research — •Gregor Hehl¹, Margot Grandl², Alexander Kovalev¹, Markus Peer², Gerhard Liebisch², Gerd Schmitz², and Andreas Volkmer¹ — ¹3rd Institute of Physics, University of Stuttgart, 70569 Stuttgart, Germany — ²University Hospital Regensburg, 93053 Regensburg, Germany

A series of technological advances have made coherent Raman scattering (CRS) microscopy a highly sensitive and chemically selective imaging tool for the noninvasive analysis of biological cells. To facilitate a spectral identification and quantitative analysis of intracellular lipid species of a priori unknown composition by CRS microscopy, we developed the technique of hyperspectral coherent anti-Stokes Raman scattering (CARS) imaging. Here, we report on its exemplifying applications in lipidomic research. We investigated the uptake mechanism of atherogenic model lipoproteins into THP-1 macrophages. We observed a decrease of the degree of acyl unsaturation within individual lipid droplets in those cells, which have been loaded with oxidized lipoproteins, compared to the unperturbed cell. Furthermore, we addressed the question: Can an intracellular Raman-based spectroscopy selectively identify and detect minor molecular structure differences between the different lipid oxidation pathways in the living cell? Corresponding mass spectra of the respective cellular lipid contents were compared and correlated with our results obtained from Raman and CRS microscopy, and will also be presented.