Live imaging of stem cells in the germarium of the Drosophila ovary using a reusable gas-permeable imaging chamber View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2018-11

AUTHORS

Amy Reilein, Elisa Cimetta, Nina M. Tandon, Daniel Kalderon, Gordana Vunjak-Novakovic

ABSTRACT

Live imaging of stem cells and their support cells can be used to visualize cellular dynamics and fluctuations of intracellular signals, proteins, and organelles in order to better understand stem cell behavior in the niche. We describe a simple protocol for imaging stem cells in the Drosophila ovary that improves on alternative protocols in that flies of any age can be used, dissection is simplified because the epithelial sheath that surrounds each ovariole need not be removed, and ovarioles are imaged in a closed chamber with a large volume of medium that buffers oxygen, pH, and temperature. We also describe how to construct the imaging chamber, which can be easily modified and used to image other tissues and non-adherent cells. Imaging is limited by follicle cells moving out of the germarium in culture around the time of egg chamber budding; however, the epithelial sheath delays this abnormal cell migration. This protocol requires an hour to prepare the ovarioles, followed by half an hour on the confocal microscope to locate germaria and set z limits. Successful imaging time depends on germarial morphology at the time of dissection, but we suggest 10-11 h to encompass all specimens. More... »

PAGES

2601-2614

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/s41596-018-0054-1

DOI

http://dx.doi.org/10.1038/s41596-018-0054-1

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1107763485

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/30349048


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