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J Microsc. 2012 Jun;246(3):237-247. doi: 10.1111/j.1365-2818.2012.03613.x.

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

Author information

1
Laboratory of Cardiac Energetics National Heart Lung and Blood Institute, Princeton, New Jersey, USA.
2
CIT Signal Processing Group, Princeton, New Jersey, USA.
3
Siemens Corporate Research, Princeton, New Jersey, USA.
#
Contributed equally

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.

PMID:
22582797
PMCID:
PMC3799900
DOI:
10.1111/j.1365-2818.2012.03613.x
[Indexed for MEDLINE]
Free PMC Article

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