Dynamic light sheet generation and fluorescence imaging behind turbid media

Dynamic light sheet generation and fluorescence imaging behind turbid media Background: Light sheet microscopy became a popular tool allowing fast imaging with reduced out of focus light. However, when light penetrates turbid media such as biological tissues, multiple scattering scrambles the illumination into a speckle pattern and severely challenges conventional fluorescence imaging with focused light or with a light sheet. In this article, we present generation of light sheet type illumination patterns despite scattering. Methods: We optimize the wave-front of the incoming light to transform the speckle pattern behind the scattering layer into a light sheet within the region of interest. We utilize a fast spatial light modulator for phase modulation and a genetic optimization algorithm. The light pattern behind the scattering layer is detected via a clear detection path and acts as a feedback signal for the algorithm. Results: We enabled homogenous light sheet illumination behind turbid media and enhanced the signal of fluorescent beads selectively at the desired focal plane up to eight times on average. The technique is capable to compensate the dynamic changes of the speckle pattern as well, as shown on samples consisting of living drosophila pupae. Conclusion: Our technique shows that not only single foci, but also a homogenous light sheet illumination can http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of the European Optical Society-Rapid Publications Springer Journals

Dynamic light sheet generation and fluorescence imaging behind turbid media

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Publisher
Springer Journals
Copyright
Copyright © 2018 by The Author(s)
Subject
Physics; Classical Electrodynamics
eISSN
1990-2573
D.O.I.
10.1186/s41476-018-0074-z
Publisher site
See Article on Publisher Site

Abstract

Background: Light sheet microscopy became a popular tool allowing fast imaging with reduced out of focus light. However, when light penetrates turbid media such as biological tissues, multiple scattering scrambles the illumination into a speckle pattern and severely challenges conventional fluorescence imaging with focused light or with a light sheet. In this article, we present generation of light sheet type illumination patterns despite scattering. Methods: We optimize the wave-front of the incoming light to transform the speckle pattern behind the scattering layer into a light sheet within the region of interest. We utilize a fast spatial light modulator for phase modulation and a genetic optimization algorithm. The light pattern behind the scattering layer is detected via a clear detection path and acts as a feedback signal for the algorithm. Results: We enabled homogenous light sheet illumination behind turbid media and enhanced the signal of fluorescent beads selectively at the desired focal plane up to eight times on average. The technique is capable to compensate the dynamic changes of the speckle pattern as well, as shown on samples consisting of living drosophila pupae. Conclusion: Our technique shows that not only single foci, but also a homogenous light sheet illumination can

Journal

Journal of the European Optical Society-Rapid PublicationsSpringer Journals

Published: Feb 21, 2018

References

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