Structural cell biology with smart STED microscopy
Laurell F. Kessler (Goethe University)
Enya S. Berrevoets (TU Delft - Applied Sciences)
Ashwin Balakrishnan (Goethe University)
Ellen K. Okuda (Goethe University)
Michaela Müller-Mcnicoll (Goethe University)
Bernd Rieger (TU Delft - Applied Sciences, TU Delft - ImPhys/Computational Imaging)
Sjoerd Stallinga (TU Delft - Applied Sciences)
Mike Heilemann (Goethe University)
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Abstract
We developed a fully-automated smart fluorescence microscopy workflow that combines fast confocal microscopy, morphological object screening, event-triggered and targeted 3D super-resolution STED microscopy and quantitative image analysis. This workflow was tailored to search for and identify rare objects or events in cells, and to subsequently direct the full performance of the microscope towards a nanoscale characterization of these objects. Using this workflow, we measured the 3D nano-morphology of paraspeckles, a phase-separated membrane-less organelle (MLO) located in the nucleus of eukaryotic cells. Furthermore, we applied this workflow to detect cell organelle contact sites in living cells. This smart microscopy approach is resource-efficient, enables targeted and high-throughput characterization of rare objects and events, and is transferable to a large variety of cellular structures.