| First Authors | Ivana Viktorinová |
|---|---|
| Authors | Ivana Viktorinová, Robert Haase, Tobias Pietzsch, Ian Henry, Pavel Tomancak |
| Corresponding Authors | Ivana Viktorinová |
| Last Authors | Pavel Tomancak |
| Journal Name | Journal of visualized experiments : JoVE (J Vis Exp) |
| Volume | 148 |
| Issue | |
| Article Number | e58587 |
| Open Access | false |
| Print Publication Date | 2019-06-03 |
| Online Publication Date | 2019-06-03 |
| Abstract | Drosophila immature eggs are called egg chambers, and their structure resembles primitive organs that undergo morphological changes from a round to an ellipsoid shape during development. This developmental process is called oogenesis and is crucial to generating functional mature eggs to secure the next fly generation. For these reasons, egg chambers have served as an ideal and relevant model to understand animal organ development. Several in vitro culturing protocols have been developed, but there are several disadvantages to these protocols. One involves the application of various covers that exert an artificial pressure on the imaged egg chambers in order to immobilize them and to increase the imaged acquisition plane of the circumferential surface of the analyzed egg chambers. Such an approach may negatively influence the behavior of the thin actomyosin machinery that generates the power to rotate egg chambers around their longer axis. Thus, to overcome this limitation, we culture Drosophila egg chambers freely in the media in order to reliably analyze actomyosin machinery along the circumference of egg chambers. In the first part of the protocol, we provide a manual detailing how to analyze the actomyosin machinery in a limited acquisition plane at the local cellular scale (up to 15 cells). In the second part of the protocol, we provide users with a new Fiji-based plugin that allows the simple extraction of a defined thin layer of the egg chambers' circumferential surface. The following protocol then describes how to analyze actomyosin signals at the tissue scale (>50 cells). Finally, we pinpoint the limitations of these approaches at both the local cellular and tissue scales and discuss its potential future development and possible applications. |
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| Affiliated With | Postdoc first author, Postdoc first female, Postdocs, Scientific Computing Facility, Tomancak, CSBD |
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| Publication Status | Published |
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| DOI | 10.3791/58587 |
| PubMed ID | 31205315 |
| WebOfScience Link | WOS:000473295900009 |
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| Display Publisher Download Only | false |
| Visible On MPI-CBG Website | true |
| PDF Downloadable | true |
| Created By | thuem |
| Added Date | 2019-06-07 |
| Last Edited By | herbst |
| Last Edited Date | 2021-05-10 18:31:21.18 |
| Library ID | 7407 |
| Document ID | |
| Entry Complete | true |
| eDoc Compliant | true |
| Include in Edoc Report | true |
| In Pure | false |
| Ready for eDoc Export | false |
| Author Affiliations Complete | true |
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