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Functional Imaging with Reinforcement, Eyetracking, and Physiological Monitoring
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Removing physiological motion from intravital and clinical functional imaging data.

Sean C Warren1,2, Max Nobis1,2, Astrid Magenau1,2

  • 1Kinghorn Cancer Centre, Garvan Institute of Medical Research, University of New South Wales, Sydney, Australia.

Elife
|July 10, 2018
PubMed
Summary

Galene software corrects physiological motion blur in microscopy images. This enables quantitative analysis of dynamic biological processes using fluorescence lifetime imaging (FLIM) even in challenging in vivo conditions.

Keywords:
FLIMFRETcell biologycomputational biologyhumanintravital microscopymotion correctionmousemultiphotonsystems biology

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Area of Science:

  • Biomedical imaging
  • Microscopy
  • Quantitative biology

Background:

  • Intravital microscopy offers insights into biological processes in their native environment.
  • Physiological motion (respiration, heartbeat, peristalsis) significantly challenges quantitative imaging, especially fluorescence lifetime imaging (FLIM).

Purpose of the Study:

  • To present and benchmark Galene, a software tool for correcting motion blur in microscopy data.
  • To enable quantitative FLIM imaging in the presence of physiological motion.

Main Methods:

  • Developed Galene, a multi-platform software for image-based motion correction.
  • Applied Galene to 2D and 3D laser scanning fluorescence microscopy data, including time-resolved data.
  • Validated Galene using intravital FLIM-FRET imaging of murine intra-abdominal organs and NADH autofluorescence of human dermal tissue.

Main Results:

  • Galene effectively corrects motion blurring caused by physiological movements.
  • Resolved intravital FLIM-FRET images of intra-abdominal organs in murine models.
  • Successfully imaged NADH autofluorescence in human dermal tissue under physiological motion.

Conclusions:

  • Galene enables FLIM imaging in situations where stable platforms are not feasible.
  • The software can recover valuable quantitative imaging data previously lost due to motion artifacts.
  • Galene expands the applicability of quantitative fluorescence microscopy in dynamic biological systems.