Active Axial Motion Compensation in Multiphoton-Excited Fluorescence Microscopy

Fuente: arXiv
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Main Authors: Kunisch, Manuel, Beutler, Sascha, Pilger, Christian, Kiefer, Friedemann, Huser, Thomas, Wirth, Benedikt
Format: Preprint
Published: 2024
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author Kunisch, Manuel
Beutler, Sascha
Pilger, Christian
Kiefer, Friedemann
Huser, Thomas
Wirth, Benedikt
author_facet Kunisch, Manuel
Beutler, Sascha
Pilger, Christian
Kiefer, Friedemann
Huser, Thomas
Wirth, Benedikt
contents In living organisms, the natural motion caused by the heartbeat, breathing, or muscle movements leads to the deformation of tissue caused by translation and stretching of the tissue structure. This effect results in the displacement or deformation of the plane of observation for intravital microscopy and causes motion-induced aberrations of the resulting image data. This, in turn, places severe limitations on the time during which specific events can be observed in intravital imaging experiments. These limitations can be overcome if the tissue motion can be compensated such that the plane of observation remains steady. We have developed a mathematical shape space model that can predict the periodic motion of a cylindrical tissue phantom resembling blood vessels. This model is then used to rapidly calculate the future position of the plane of observation of a confocal multiphoton fluorescence microscope. The focal plane is continuously adjusted to the calculated position with a piezo-actuated objective lens holder. We demonstrate active motion compensation for non-harmonic axial displacements of the vessel phantom with a field of view up to 400 $μ$m $\times$ 400 $μ$m, vertical amplitudes of more than 100 $μ$m, and at a rate of 0.5 Hz.
format Preprint
id arxiv_https___arxiv_org_abs_2401_10598
institution arXiv
publishDate 2024
record_format arxiv
spellingShingle Active Axial Motion Compensation in Multiphoton-Excited Fluorescence Microscopy
Kunisch, Manuel
Beutler, Sascha
Pilger, Christian
Kiefer, Friedemann
Huser, Thomas
Wirth, Benedikt
Optics
Image and Video Processing
Biological Physics
In living organisms, the natural motion caused by the heartbeat, breathing, or muscle movements leads to the deformation of tissue caused by translation and stretching of the tissue structure. This effect results in the displacement or deformation of the plane of observation for intravital microscopy and causes motion-induced aberrations of the resulting image data. This, in turn, places severe limitations on the time during which specific events can be observed in intravital imaging experiments. These limitations can be overcome if the tissue motion can be compensated such that the plane of observation remains steady. We have developed a mathematical shape space model that can predict the periodic motion of a cylindrical tissue phantom resembling blood vessels. This model is then used to rapidly calculate the future position of the plane of observation of a confocal multiphoton fluorescence microscope. The focal plane is continuously adjusted to the calculated position with a piezo-actuated objective lens holder. We demonstrate active motion compensation for non-harmonic axial displacements of the vessel phantom with a field of view up to 400 $μ$m $\times$ 400 $μ$m, vertical amplitudes of more than 100 $μ$m, and at a rate of 0.5 Hz.
title Active Axial Motion Compensation in Multiphoton-Excited Fluorescence Microscopy
topic Optics
Image and Video Processing
Biological Physics
url https://arxiv.org/abs/2401.10598