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BP: Fachverband Biologische Physik
BP 11: Cell Mechanics I
BP 11.5: Talk
Tuesday, March 19, 2024, 11:00–11:15, H 2032
In-situ high-throughput analysis of mitochondrial membrane tension under pathophysiological conditions — •Eric Sündermann, Bob Fregin, Jan Maurice Wilder, Doreen Biedenweg, Stefanie Spiegler, and Oliver Otto — Institute of Physics, University of Greifswald, Greifswald, Germany
The development of high-throughput methods for cell mechanical research is becoming increasingly important in biology, medicine and physics as the analysis of large samples increases the statistical robustness to identify rare cell populations and to transfer results from basic science into clinical applications. While most studies focus on 2D/3D cellular systems, little is known about how chemical and physical stress propagates inside the cell and impacts the mechanical properties of organelles.
Here, we applied membrane tension cytometry (MTC), a technology recently developed in our lab, to study the intracellular response of mitochondrial mechanics to hydrodynamic and oxidative stress. As a model system, HL60 cells have been chosen that were incubated with hydrogen peroxide to generate mitochondrial superoxide as reactive oxygen species. After staining cells with Mito Flipper-TR we took advantage of the fact that its fluorescence lifetime is proportional to the membrane tension of mitochondria. Preliminary experiments using MTC show that hydrodynamic stress propagates linearly to mitochondria inside the cytosol and that oxidative stress leads to their softening - in agreement with earlier results studying isolated mitochondria using real-time deformability cytometry.
Keywords: Fluorescence lifetime; Real-time deformability cytometry; Mitochondria mechanics; Organelle mechanics; Membrane tension cytometry