Three-dimensional motion tracking for high-resolution optical microscopy, in vivo

Laboratory of Cardiac Energetics, National Heart Lung and Blood Institute.
Journal of Microscopy (Impact Factor: 2.33). 06/2012; 246(3):237-47. DOI: 10.1111/j.1365-2818.2012.03613.x
Source: PubMed


When conducting optical imaging experiments, in vivo, the signal to noise ratio and effective spatial and temporal resolution is fundamentally limited by physiological motion of the tissue. A three-dimensional (3D) motion tracking scheme, using a multiphoton excitation microscope with a resonant galvanometer, (512 × 512 pixels at 33 frames s(-1)) is described to overcome physiological motion, in vivo. The use of commercially available graphical processing units permitted the rapid 3D cross-correlation of sequential volumes to detect displacements and adjust tissue position to track motions in near real-time. Motion phantom tests maintained micron resolution with displacement velocities of up to 200 μm min(-1), well within the drift observed in many biological tissues under physiologically relevant conditions. In vivo experiments on mouse skeletal muscle using the capillary vasculature with luminal dye as a displacement reference revealed an effective and robust method of tracking tissue motion to enable (1) signal averaging over time without compromising resolution, and (2) tracking of cellular regions during a physiological perturbation.

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    • "ization for all phases of the cardiac cycle is through active motion compensation . In this method the relative residual motion present between the imaging device ( i . e . , objective ) and the imaged tissue can be actively canceled by tracking in real time the tissue position ( Nakamura et al . , 2001 ; Lee et al . , 2008 ; Yuen et al . , 2009 ; Bakalar et al . , 2012 ) and accordingly shifting the objective . This cancelation of relative movement virtually freezes in space both the objective and imaged sample leading to motion artifact free images ."
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    Frontiers in Physiology 05/2015; 6. DOI:10.3389/fphys.2015.00147 · 3.53 Impact Factor
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    • "It is therefore recommended, whenever possible, to adopt passive mechanical stabilizers in combination with their use in a synergistic effort aimed at reducing the total amount of motion [8], [24]. Alternatively motion compensation along the plane perpendicular to the optical axis has been demonstrated by controlling an XY translational stage [12] while Z motion compensation has also been achieved using an objective focus motor [25] "
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