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Title: | Live Imaging in Planarians: Immobilization and Real-Time Visualization of Reactive Oxygen Species | Authors: | JAENEN, Vincent BIJNENS, Karolien HELEVEN, Martijn ARTOIS, Tom SMEETS, Karen |
Issue Date: | 2023 | Publisher: | Springer Protocols | Source: | Gentile, Luca (Ed.). Schmidtea mediterranea: Methods and Protocols, Springer Protocols, p. 209 -229 | Series/Report: | Methods in Molecular Biology | Series/Report no.: | 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 mapping 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 immobili-zation 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. | Keywords: | In vivo studies;Live imaging;Immobilization;Planarians;Reactive oxygen species;ROS;Developmental dynamics;Autofluorescence | Document URI: | http://hdl.handle.net/1942/41794 | ISBN: | 9781071632741 9781071632758 |
ISSN: | 1064-3745 | DOI: | 10.1007/978-1-0716-3275-8_13 | Category: | B2 | Type: | Book Section |
Appears in Collections: | Research publications |
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Chapter 13_978-1-0716-3275-8.pdf Restricted Access | Published version | 1.03 MB | Adobe PDF | View/Open Request a copy |
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