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BP: Fachverband Biologische Physik

BP 8: Posters - Bioimaging and Spectroscopy

BP 8.4: Poster

Montag, 20. März 2017, 17:30–19:30, P3

An integrated platform for rapid semi-confocal imaging and spatially resolved fluctuation microscopy — •Adal Sabri, Andreas Veres, and Matthias Weiß — Experimental Physics 1, University of Bayreuth

Fluorescence imaging is a key method to study the dynamics of biological specimen. Due to the common trade-off between spatial and temporal resolution, rapid high-quality data acquisition often comes at the cost of complex and technically challenging methods.

We report on a technique that increases the temporal resolution of image acquisition by more than an order of magnitude as compared to standard confocal microscopy approaches. Large areas (up to 450µm edge length) can be imaged rapidly with a resolution close to the diffraction limit. To this end, multiple cylindrical lenses shape a thin light sheet so that effectively only a line within a thin specimen (oriented perpendicular to the optical axis) is illuminated. This line of illumination is scanned in one spatial direction with a Galvo mirror. A slit aperture in the detection path yields an axial discrimination, thus creating a semi-confocal setup.

Swift switching to a second excitation/detection path allows for alternating between advanced rapid image acquisition and two-point fluctuation spectroscopy on smaller scales. The setup allows one to correlate fluorescence fluctuations at two selectable, spatially separated foci over time to determine local transport coefficients, hence supports the combination of a rapid imaging and the analysis of dynamic intracellular events on a subcellular scale.