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

Journal of Microscopy
M BakalarRobert S Balaban

Abstract

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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Citations

Mar 29, 2014·Journal of Microscopy·B Lucotte, R S Balaban
Oct 4, 2014·Microcirculation : the Official Journal of the Microcirculatory Society, Inc·Brian GlancyRobert S Balaban
Nov 21, 2013·Journal of Microscopy·Christian A CombsRobert S Balaban
Nov 26, 2013·IEEE Journal of Selected Topics in Quantum Electronics : a Publication of the IEEE Lasers and Electro-optics Society·Claudio VinegoniRalph Weissleder
Aug 1, 2015·Nature·Brian GlancyRobert S Balaban
Jun 2, 2015·Frontiers in Physiology·Claudio VinegoniRalph Weissleder
Mar 3, 2017·Journal of Biomedical Optics·Sungon LeeClaudio Vinegoni
Jul 3, 2015·Biomedical Optics Express·Ben SherlockChris Dunsby

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