Dresden 2009 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 11: Novel Methods
BP 11.1: Vortrag
Dienstag, 24. März 2009, 14:30–14:45, ZEU 260
Fast Dynamics of Cellular Signals Studied With a Novel Miniaturized Multi-Channel Perfusion System — •Claus Fütterer1, Luru Dai2, Hans Fried2, Arne Brombas2, Arnd Baumann2, Thomas Gensch2, and Frank Müller2 — 1Forschungszentrum Jülich, IBN-4, 52425 Jülich — 2Forschungszentrum Jülich, INB-1, 52425 Jülich
In order to study the dynamics of cellular signaling in single cells and ex vivo-tissues, we developed a novel miniaturized fluidic system that allows unparalleled fast and artefact-free solution exchange using multiple inflow channels. To this end, methods of microfabrication and numerical simulations have been involved.
HEK-293 cells were used that expressed octopamine receptors as well as the genetically encoded calcium sensor TN-L15, designed for calcium dependent Fluorescence Resonance Energy Transfer. Switching from octopamine free to octopamine containing solution leads to oscillations of the intracellular calcium concentrationof which the onset and fading out of these oscillations can be controlled with unparalleled temporal resolution.
Further we studied calcium signals in flatmounted retinae of transgenic mice that express TN-L15 in retinal ganglion cells. Cells were activated by switching the perfusion from normal extracellular solution to solution with high K+ concentration to enable the precise measurement of the intracellular calcium concentration. The high temporal resolution achieved in our set-up revealed differences in the response of individual ganglion cells that had not been detected before.