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    Area of Science:

    • Biomedical Engineering
    • Optical Microscopy
    • In Vivo Imaging

    Background:

    • Living organisms exhibit natural motion (heartbeat, breathing, muscle movement) causing tissue deformation.
    • This motion displaces the observation plane in intravital microscopy, leading to motion-induced aberrations and limiting observation time.
    • Current methods struggle to overcome these limitations for observing dynamic biological processes.

    Purpose of the Study:

    • To develop a method for active motion compensation in intravital microscopy.
    • To overcome limitations imposed by physiological motion on imaging dynamic events.
    • To enable stable, long-term observation of biological processes in vivo.

    Main Methods:

    • Developed a mathematical shape space model to predict periodic motion of cylindrical tissue phantoms (e.g., blood vessels).
    • Utilized the model to calculate future positions of the microscope's observation plane.
    • Implemented a piezo-actuated objective lens holder for continuous focal plane adjustment to compensate for motion.

    Main Results:

    • Demonstrated active motion compensation for non-harmonic axial displacements of a vessel phantom.
    • Successfully compensated for motion with vertical amplitudes exceeding 100 µm at 0.5 Hz.
    • Maintained a stable observation plane within a field of view up to 400 µm × 400 µm.

    Conclusions:

    • The developed mathematical model and active compensation system effectively counteract physiological tissue motion during intravital microscopy.
    • This approach significantly enhances the ability to observe dynamic biological events by stabilizing the imaging plane.
    • The method holds promise for improving the duration and quality of in vivo imaging experiments.