SKM 2023 – scientific programme
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BP: Fachverband Biologische Physik
BP 28: Poster Session II
BP 28.6: Poster
Thursday, March 30, 2023, 18:00–20:00, P2/EG
2D polarization fluorescence imaging (2DPOLIM) - Evaluation of depolarization artifacts in fixed four channel detection — •Yunhao Mei1,2, Asad Hafeez1,2, Yutong Wang1,2, Mohammad Soltaninezhad1,2, Rainer Heintzmann1,2, and Daniela Täuber1,2 — 1Heintzmann Lab, Leibniz Institute of Photonic Technology, Jena — 2Institute of Physical Chemistry & Abbe Center of Photonics, Friedrich-Schiller-University Jena, Germany
Fluorescence polarization and anisotropy measurements are widely used in Diagnostics and Imaging in the Life Sciences. Most common is the use of linearly polarized excitation together with two-channel detection of linear polarization parallel and perpendicular to the polarization in excitation. However, when applied to anisotropic samples, the result may depend on the lab frame in respect to structural alignments within the sample, which can be preventend by collecting the sample fluorescence in more polarization angles[1]. We designed a four-channel detection for evaluating Förster Resonance Energy Transfer between similar fluorescence labels, the so-called homo-FRET or emFRET. The quantitative discrimination of FRET via tiny changes in polarization resolved fluorescence intensities requires highly sensitive cameras and well-designed and calibrated detection channels. Wire grids provide high-quality polarization properties when used in normal incidence in transmission but may introduce depolarization when implemented tilted in respect to the optical axis. We studied several possible arrangements and compared the introduced depolarizations. [1] R. Camacho et al. Adv. Mater. 2019, 1805671.