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<meta name="description" content="An IDEAS project for improving commercial microscopy system that adjusts to and compensates for motion when studying in vivo subcellular structures." />
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<title>Two-Photon Excitation Fluorescence Microscopy Motion Tracking to Study in-vivo Subcellular Structures | National Institute of Biomedical Imaging and Bioengineering</title>
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<span class="a2a_kit a2a_kit_size_32 addtoany_list" data-a2a-url="https://www.nibib.nih.gov/labs-at-nibib/ideas-lab/projects/two-photon-excitation-fluorescence-microscopy-motion-tracking-study-vivo-subcellular-structures" data-a2a-title="Two-Photon Excitation Fluorescence Microscopy Motion Tracking to Study in-vivo Subcellular Structures"><a class="a2a_button_email"><span class="a2a_svg a2a_s_a2a a2a_s_email a2a-box"><img src="/themes/custom/nibib/assets/img/email-a2a.svg" border="0" alt="Facebook" width="27" height="27"></span></a><a class="a2a_button_facebook"><span class="a2a_svg a2a_s_a2a a2a_s_facebook a2a-box"><img src="/themes/custom/nibib/assets/img/facebook-a2a.svg" border="0" alt="Facebook" width="27" height="27"></span></a><a class="a2a_button_linkedin"><span class="a2a_svg a2a_s_a2a a2a_s_linkedin a2a-box"><img src="/themes/custom/nibib/assets/img/linkedin-a2a.svg" border="0" alt="Facebook" width="27" height="27"></span></a><a class="a2a_button_twitter"><span class="a2a_svg a2a_s_a2a a2a_s_x a2a-box"><img src="/themes/custom/nibib/assets/img/x-a2a.svg" border="0" alt="Facebook" width="27" height="27"></span
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<h1 class="margin-0 page-header">
<span>Two-Photon Excitation Fluorescence Microscopy Motion Tracking to Study in-vivo Subcellular Structures</span>
</h1>
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<article data-history-node-id="136406" about="/labs-at-nibib/ideas-lab/projects/two-photon-excitation-fluorescence-microscopy-motion-tracking-study-vivo-subcellular-structures" class="ideas-projects node--ideas-projects node--type-ideas-projects">
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<div class="field field--name-field-paragraphs field--type-entity-reference-revisions field--label-above">
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<h2 class="field--label">Collaborators</h2>
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<div class="field field--name-field-collaborator field--type-link field--label-hidden field__item"><span>Robert Balaban, Ph.D.</span></div>
<div class="field field--name-field-institution field--type-string field--label-hidden field__item">Laboratory of Cardiac Energetics, NHLBI</div>
</div>
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<span class="grid-col flex-auto"> <div class="paragraph paragraph--type--collaborators paragraph--view-mode--default">
<div class="field field--name-field-collaborator field--type-link field--label-hidden field__item"><span>David Chess, Ph.D.</span></div>
<div class="field field--name-field-institution field--type-string field--label-hidden field__item">Laboratory of Cardiac Energetics, NHLBI</div>
</div>
</span>
<span class="grid-col flex-auto"> <div class="paragraph paragraph--type--collaborators paragraph--view-mode--default">
<div class="field field--name-field-collaborator field--type-link field--label-hidden field__item"><span>James Schroeder, Ph.D.</span></div>
<div class="field field--name-field-institution field--type-string field--label-hidden field__item">Laboratory of Cardiac Energetics, NHLBI</div>
</div>
</span>
<span class="grid-col flex-auto"> <div class="paragraph paragraph--type--collaborators paragraph--view-mode--default">
<div class="field field--name-field-collaborator field--type-link field--label-hidden field__item"><span>Matthew Bakalar</span></div>
<div class="field field--name-field-institution field--type-string field--label-hidden field__item">Laboratory of Cardiac Energetics, NHLBI (formerly with IDEAS)</div>
</div>
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<span class="grid-col flex-auto"> <div class="paragraph paragraph--type--collaborators paragraph--view-mode--default">
<div class="field field--name-field-collaborator field--type-link field--label-hidden field__item"><span>Merav Luger-Hamer, Ph.D.</span></div>
<div class="field field--name-field-institution field--type-string field--label-hidden field__item">Laboratory of Cardiac Energetics, NHLBI</div>
</div>
</span>
<span class="grid-col flex-auto"> <div class="paragraph paragraph--type--collaborators paragraph--view-mode--default">
<div class="field field--name-field-collaborator field--type-link field--label-hidden field__item"><span>Peter Kellman, Ph.D.</span></div>
<div class="field field--name-field-institution field--type-string field--label-hidden field__item">Laboratory of Cardiac Energetics, NHLBI</div>
</div>
</span>
<span class="grid-col flex-auto"> <div class="paragraph paragraph--type--collaborators paragraph--view-mode--default">
<div class="field field--name-field-collaborator field--type-link field--label-hidden field__item"><span>Brian Glancy, Ph.D.</span></div>
<div class="field field--name-field-institution field--type-string field--label-hidden field__item">Laboratory of Cardiac Energetics, NHLBI</div>
</div>
</span>
<span class="grid-col flex-auto"> <div class="paragraph paragraph--type--collaborators paragraph--view-mode--default">
<div class="field field--name-field-collaborator field--type-link field--label-hidden field__item"><span>Michael Hansen, Ph.D.</span></div>
<div class="field field--name-field-institution field--type-string field--label-hidden field__item">Laboratory of Cardiac Energetics, NHLBI</div>
</div>
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<span class="grid-col flex-auto"> <div class="paragraph paragraph--type--collaborators paragraph--view-mode--default">
<div class="field field--name-field-collaborator field--type-link field--label-hidden field__item"><span>Bertrand Lucotte, Ph.D.</span></div>
<div class="field field--name-field-institution field--type-string field--label-hidden field__item">Laboratory of Cardiac Energetics, NHLBI</div>
</div>
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<div class="ideas-project-section-title">
<h2>Project Brief</h2>
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<div class="field field--name-body field--type-text-with-summary field--label-hidden field__item"><p>IDEAS collaborated with the NHLBI Laboratory of Cardiac Energetics to develop a methodology enabling the study in vivo sub-cellular structures and signaling processes in real-time. &nbsp;Multi-photon fluorescence imaging provides improved tissue penetration, sensitivity, and information content when investigating dynamic intercellular events within living tissue, but physiological motion degrades the quality of these images and makes temporal observations challenging. &nbsp;Our system implementation, designed to be functional in tandem with the commercial two-photon microscope system, adjusts the field-of-view and focal plane of the microscope in real-time to compensate for the natural physiological motion of living tissue. &nbsp;The function and performance of the commercial system is unaffected. &nbsp;The motion tracking system intercepts the images as they are generated by the microscope, creating a multi-slice volume. &nbsp;The system utilizes a high-performance GPU platform to correlate each real-time volume to an initial reference volume to quantify the amount and direction of tissue motion. &nbsp;This motion information is used to adjust the microscope XY stage and objective to dynamically correct for the tissue motion.&nbsp; IDEAS&nbsp;integrated the custom hardware and GPU, and developed the custom software for this system.&nbsp;</p>
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<p>&nbsp;</p>
<figure role="group" class="align-center">
<img alt="Time-lapse volume of with and without motion tracking correction" data-entity-type="file" data-entity-uuid="f636001a-eb8b-4833-be82-016fdcb2e74f" src="/sites/default/files/inline-images/Twophoton3.png" width="640" height="238" loading="lazy"></p>
<figcaption>Time-lapse volume of with and without motion tracking correction</figcaption>
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<div class="field field--name-field-card-text field--type-text-long field--label-hidden field__item"><p><a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3799900/">Three Dimensional Motion Tracking for High Resolution Optical Microscopy, in vivo</a></p>
<p><a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3115409/">Rapid Overlapping-Volume Acquisition and Reconstruction (ROVAR): Automated 3D Tiling for High-Resolution, Large Field-of-View Optical Microscopy</a></p>
<p><a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3209509/">Subcellular Motion Compensation for Minimally Invasive Microscopy, In Vivo. Evidence for Oxygen Gradients in Resting Muscle</a></p>
<p>Motion Tracking Using Optical Navigation During in-vivo Two-Photon Microscopy&nbsp;<em>(unavailable)</em></p>
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