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Live Imaging in Planarians: Immobilization and Real-Time Visualization of Reactive Oxygen Species

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Schmidtea Mediterranea

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2680))

Abstract

Imaging of living animals allows the study of metabolic processes in relation to cellular structures or larger functional entities. To enable in vivo imaging during long-term time-lapses in planarians, we combined and optimized existing protocols, resulting in an easily reproducible and inexpensive procedure. Immobilization with low-melting-point agarose eliminates the use of anesthetics, avoids interfering with the animal during imaging—functionally or physically—and allows recovering the organisms after the imaging procedure. As an example, we used the immobilization workflow to image the highly dynamic and fast-changing reactive oxygen species (ROS) in living animals. These reactive signaling molecules can only be studied in vivo and map** their location and dynamics during different physiological conditions is crucial to understand their role in developmental processes and regeneration. In the current protocol, we describe both the immobilization and ROS detection procedure. We used the intensity of the signals together with pharmacological inhibitors to validate the signal specificity and to distinguish it from the autofluorescent nature of the planarian.

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Authors and Affiliations

  1. Centre for Environmental Sciences, Hasselt University, Diepenbeek, Belgium

    Vincent Jaenen, Karolien Bijnens, Martijn Heleven, Tom Artois & Karen Smeets

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  1. Vincent Jaenen
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  2. Karolien Bijnens
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  3. Martijn Heleven
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Correspondence to Karen Smeets .

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  1. Department of Animal Physiology, University of Osnabrück, Osnabrück, Germany

    Luca Gentile

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Jaenen, V., Bijnens, K., Heleven, M., Artois, T., Smeets, K. (2023). Live Imaging in Planarians: Immobilization and Real-Time Visualization of Reactive Oxygen Species. In: Gentile, L. (eds) Schmidtea Mediterranea. Methods in Molecular Biology, vol 2680. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3275-8_13

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  • DOI: https://doi.org/10.1007/978-1-0716-3275-8_13

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