Three-dimensional motion tracking for high-resolution optical microscopy, in vivo.
J Microsc
; 246(3): 237-247, 2012 Jun.
Article
em En
| MEDLINE
| ID: mdl-22582797
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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Base de dados:
MEDLINE
Assunto principal:
Músculo Esquelético
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Microscopia de Vídeo
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Imageamento Tridimensional
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Locomoção
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Microscopia de Fluorescência
Limite:
Animals
Idioma:
En
Ano de publicação:
2012
Tipo de documento:
Article