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Dynamic light sheet generation and fluorescence imaging behind turbid media.

Jale Schneider1, Christof M Aegerter1

  • 1Physik-Institut, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.

Journal of the European Optical Society. Rapid Publications
|March 24, 2018
PubMed
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We developed a method to create light sheet illumination through scattering biological tissues. This technique optimizes light wavefronts to overcome scattering, enabling clearer imaging in turbid media.

Keywords:
Imaging through turbid mediaLight sheet microscopyPhase modulationWave-front shaping

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Area of Science:

  • Biophotonics
  • Microscopy

Background:

  • Light sheet microscopy offers fast imaging with reduced out-of-focus light.
  • Scattering in turbid media like biological tissues creates speckle patterns, challenging conventional imaging.
  • Generating structured illumination through scattering media is a significant hurdle in biological imaging.

Purpose of the Study:

  • To generate light sheet illumination patterns through scattering media.
  • To overcome the limitations of light penetration in turbid biological samples.
  • To improve fluorescence imaging in scattering environments.

Main Methods:

  • Optimized light wavefronts using a spatial light modulator and genetic algorithm.
  • Transformed speckle patterns behind scattering layers into light sheets.
  • Utilized a clear detection path for feedback to the optimization algorithm.

Main Results:

  • Achieved homogenous light sheet illumination behind turbid media.
  • Enhanced fluorescent bead signal by up to eight times at the focal plane.
  • Demonstrated compensation for dynamic speckle pattern changes in living Drosophila pupae.

Conclusions:

  • Homogenous light sheet illumination can be created and maintained behind static and dynamic scattering media.
  • The technique enhances imaging capabilities in challenging biological settings.
  • A fluorescent probe can provide feedback for turbid detection paths, increasing applicability.