Berlin 2018 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 23: Bioimaging and Biopspectroscopy II
BP 23.4: Vortrag
Mittwoch, 14. März 2018, 15:45–16:00, H 2013
Characterisation of Metabolic Dynamics by Fluorescence Lifetime Imaging Microscopy of NAD(P)H — •André Weber1, Yury Prokazov1, Marcus Hauser2, and Werner Zuschratter1 — 1Leibniz-Institut für Neurobiologie Magdeburg, Germany — 2Institut für Biometrie und Medizinische Informatik, Otto-von-Guericke- Universität Magdeburg, Germany
The energy metabolism of eucaryotic cells can show complex dynamics, i.e. glycolytic oscillations. Monitoring this intrinsic behaviour by fluorescence microscopy is influence the metabolism, which is sensitive to excitation light intensities, especially in UV range.
We show a low light imaging approach using a single photon counting position sensitive detector working with laser intensities below 3mW/cm2 and a time resolution below 90ps. For excitation of intracellular NAD(P)H a 8 MHz pulsed frequency-tripled Nd:vanadate laser tuned at 355 nm was used. The analysis of the complex fluorescence decay of NAD(P)H in intact yeast cells revealed 4 molecular species with characteristic fluorescence lifetimes showing individual behaviour and glycolytic oscillations as response to glucose addition. Laser intensities higher 3mW/cm2 did not lead to long lasting glycolytic oscillations.