Dresden 2009 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 11: Novel Methods
BP 11.4: Vortrag
Dienstag, 24. März 2009, 15:15–15:30, ZEU 260
Label-free bioimaging of living human glioblastoma cells by confocal Raman microscopy — •Alexander M. Gigler1, Katharina Klein2, Guido Piontek2, Thomas Aschenbrenner3, Wolfram Bunk3, Gregor Morfill3, Jürgen Schlegel2, and Robert W. Stark1 — 1Center for Nanoscience (CeNS) and Sect. Crystallography, LMU-München, D-80333 München — 2Neuropathology, TU-München, Klinikum rechts der Isar, D-81675 München — 3MPI for Extraterrestrial Physics, D-85748 Garching
Label-free imaging by confocal Raman spectroscopy is becoming a promising alternative to established methods for cell imaging requiring fixation and the use of fluorescent markers. With our setup we are able to image living cells at a high resolution in buffer solution (PBS). Different cellular compartments can be visualized and directly compared to immunofluorescence microscopy (IF). The comparison of Raman and IF image sets allows an assignment of organelles such as nucleus, endoplasmatic reticulum, and mitochondria. From the assigned areas we obtained average spectra of the compartments resulting in an individual spectral fingerprint for each specific region. These fingerprints can in turn be used to define spectral filters for mapping in an iterative procedure. Spectral maps of single cells provide the full set of biochemical information contained in the selected focal plane. To this end, we are using IF staining methods to verify our observations and assignments. On the long run, our aim is to identify specific molecular markers for therapeutic targeting and discriminate between cells of different lines or differentiation states based on spectral information.