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  • Artefact-free timelapse light sheet microscopy in, on and around the beating heart
  • Artefact-free timelapse light sheet microscopy in, on and around the beating heart

    Abstract number
    192
    Presentation Form
    Submitted Talk
    Corresponding Email
    [email protected]
    Session
    Stream 6 (Frontiers): Lightsheet Microscopy: Development and Application
    Authors
    Dr Jonathan Taylor (2), Carl J Nelson (2), Finnius A Bruton (1), Aryan Kaveh (1), Vytautas Zickus (2), Carl S Tucker (1), Martin A Denvir (1)
    Affiliations
    1. University of Edinburgh
    2. University of Glasgow
    Keywords

    light sheet microscopy, motion artefacts, in vivo imaging, optical gating, timelapse imaging


    Abstract text

    The heart is a particularly challenging organ to image in 3D timelapse, due to its constant motion. To image processes on timescales of minutes to hours (such as heart development, cell migration, repair and regeneration) demands some form of synchronized image acquisition in order to isolate the high-frequency heartbeat motion from the lower-speed morphological changes of interest. It is also important that any solution should avoid undesirable physiological effects on the sample. Our techniques for prospective optically-gated light sheet microscopy have given us a routine tool for day-long 3D timelapse imaging of the developing zebrafish heart, enabling us to image immune cell dynamics during the inflammatory response to cardiac injury, and revealing unexpected proliferative behaviour of cardiomyocytes during trabeculation.


    We have also been able to map out blood flow fields within the beating heart, by using related synchronization techniques to extract high-precision measurements from noisy and low-quality raw image data. Not only does this work offer a bridge between direct experimental measurements and computational fluid-structure modelling of heart development, it also points the way towards exciting possibilities in integrated studies of the developmental coupling between heart structure, fluid flow and electrical activity.


    Finally, motion artefacts have an impact on high-resolution imaging in other tissues outside the heart, and we will examine the scope for both motion-synchronized image acquisition and post-acquisition correction of motion artefacts.


    These scenarios highlight the fact that “resolution” in a microscope does not tell the whole story. We can only get the most out of the microscope if we are able to cope with motion artefacts. I will finish by considering future prospects for motion-aware microscopy, with the aim of extending such techniques to unmodified commercial microscope systems so that every biology lab can benefit from motion-stabilized in vivo imaging.

    References

    Taylor, J.M., Nelson, C.J., Bruton, F.A. et al. Adaptive prospective optical gating enables day-long 3D time-lapse imaging of the beating embryonic zebrafish heart. Nat Commun10, 5173 (2019)