Long-term in vivo imaging of Drosophila larvae.


Journal

Nature protocols
ISSN: 1750-2799
Titre abrégé: Nat Protoc
Pays: England
ID NLM: 101284307

Informations de publication

Date de publication:
03 2020
Historique:
received: 22 07 2019
accepted: 09 12 2019
pubmed: 12 2 2020
medline: 22 4 2020
entrez: 12 2 2020
Statut: ppublish

Résumé

The Drosophila larva has been used to investigate many processes in cell biology, including morphogenesis, physiology and responses to drugs and new therapeutic compounds. Despite its enormous potential as a model system, longer-term live imaging has been technically challenging because of a lack of efficient methods for immobilizing larvae for extended periods. We describe here a simple procedure for anesthetization and uninterrupted long-term in vivo imaging of the epidermis and other larval organs, including gut, imaginal discs, neurons, fat body, tracheae, muscles and hemocytes, for up to 8 h. We also include a procedure for probing cell properties by laser ablation. We provide a survey of the effects of different anesthetics, demonstrating that short exposure to diethyl ether is the most effective for long-term immobilization of larvae. This protocol does not require specific expertise beyond basic Drosophila genetics and husbandry, and confocal microscopy. It enables high-resolution studies of many systemic and subcellular processes in larvae.

Identifiants

pubmed: 32042177
doi: 10.1038/s41596-019-0282-z
pii: 10.1038/s41596-019-0282-z
doi:

Substances chimiques

Ether 0F5N573A2Y

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

1158-1187

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Auteurs

Parisa Kakanj (P)

Institute for Genetics, University of Cologne, Cologne, Germany. pkakanj@uni-koeln.de.
Department of Biological Mechanisms of Ageing, Max Planck Institute for Biology of Ageing, Cologne, Germany. pkakanj@uni-koeln.de.
Center for Molecular Medicine Cologne, University of Cologne, Cologne, Germany. pkakanj@uni-koeln.de.
Directors' Research Unit, European Molecular Biology Laboratory, Heidelberg, Germany. pkakanj@uni-koeln.de.
Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases, University of Cologne, Cologne, Germany. pkakanj@uni-koeln.de.

Sabine A Eming (SA)

Center for Molecular Medicine Cologne, University of Cologne, Cologne, Germany.
Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases, University of Cologne, Cologne, Germany.
Department of Dermatology, University of Cologne, Cologne, Germany.

Linda Partridge (L)

Department of Biological Mechanisms of Ageing, Max Planck Institute for Biology of Ageing, Cologne, Germany. Partridge@age.mpg.de.
Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases, University of Cologne, Cologne, Germany. Partridge@age.mpg.de.
Institute of Healthy Ageing, Department of Genetics, Evolution, and Environment, University College London, London, UK. Partridge@age.mpg.de.

Maria Leptin (M)

Institute for Genetics, University of Cologne, Cologne, Germany. mleptin@uni-koeln.de.
Center for Molecular Medicine Cologne, University of Cologne, Cologne, Germany. mleptin@uni-koeln.de.
Directors' Research Unit, European Molecular Biology Laboratory, Heidelberg, Germany. mleptin@uni-koeln.de.
Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases, University of Cologne, Cologne, Germany. mleptin@uni-koeln.de.

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