A live-imaging protocol for tracking intestinal stem cell divisions in the Drosophila melanogaster pupal midgut

Song Wu; Ruizhi Tang; Benjamin Ohlstein; Zheng Guo

doi:10.1016/j.xpro.2023.102749

STAR Protocols (Dec 2023)

A live-imaging protocol for tracking intestinal stem cell divisions in the Drosophila melanogaster pupal midgut

Song Wu,
Ruizhi Tang,
Benjamin Ohlstein,
Zheng Guo

Affiliations

Song Wu: Department of Pharmacology, Bioengineering and Food College, Hubei University of Technology, Wuhan 430068, China; Department of Medical Genetics, School of Basic Medicine, Institute for Brain Research, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China; Corresponding author
Ruizhi Tang: Department of Medical Genetics, School of Basic Medicine, Institute for Brain Research, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China
Benjamin Ohlstein: Children’s Research Institute and Department of Pediatrics, University of Texas Southwestern Medical Center, Dallas, TX, USA
Zheng Guo: Department of Medical Genetics, School of Basic Medicine, Institute for Brain Research, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China; Cell Architecture Research Center, Huazhong University of Science and Technology, Wuhan 430030, China; Corresponding author

DOI: https://doi.org/10.1016/j.xpro.2023.102749
Journal volume & issue: Vol. 4, no. 4
p. 102749

Abstract

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Summary: Establishing a long-term ex vivo observation of the intestinal stem cell (ISC) is crucial to help understand the formation and homeostasis of the intestinal epithelium. Here, we present a protocol for tracking the division of Drosophila pupal ISCs during pupal midgut development. We describe steps for dissecting, mounting, and live imaging the pupal midgut. We then detail procedures for fluorescence quantification of each cell. This protocol can be applied to other fluorescently tagged proteins.For complete details on the use and execution of this protocol, please refer to Wu et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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