A Fast Projection Imaging Method for the Quantification of the Dynamics of Endosome Maturation

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Poster Session 1
Xian Hu (1), Salma Jalah (5), Michael Sheetz (3), Oddmund Bakke (4), Felix Margadant (2)
1. Center for Molecular Medicine Norway, University of Oslo, Norway
2. Center of Cancel Reprogramming, Radium Hospital, Norway
3. Department of Biochemistry & Molecular Biology, UTMB, US
4. Department of Biosciences, University of Oslo, Norway
5. University College London, UK
Abstract text

Despite progress made in confocal microscopy, even fast systems still have insufficient temporal resolution for detailed live-cell volume imaging, such as tracking rapid movement of membrane vesicles in three-dimensional space. By sacrificing detailed information in the Z-direction, we propose a new imaging modality that involves capturing fast ‘projections’ from the field of depth and shortens imaging time by approximately an order of magnitude as compared to standard volumetric confocal imaging. The implementation minimally requires two synchronized control signals that drive a piezo stage and trigger the camera exposure. The device generating the signals has been tested on spinning disk confocal and instant structured-illumination-microscopy (iSIM) microscopes. Since the initial publication of the method, we have made several improvements on system stablization and stepping speed, and have implemented the system onto two other imaging platforms other than our own microscope.

  Hu, X., Jalal, S., Sheetz, M., Bakke, O., Margadant, F., 2020. Micro-stepping Extended Focus reduces photobleaching and preserves structured illumination super-resolution features. J. Cell. Sci. https://doi.org/10.1242/jcs.240796