Universität zu Köln

Kakanj, Parisa, Eming, Sabine A., Partridge, Linda ORCID: 0000-0001-9615-0094 and Leptin, Maria ORCID: 0000-0001-7097-348X (2020). Long-term in vivo imaging of Drosophila larvae. Nat. Protoc., 15 (3). S. 1158 - 1188. LONDON: NATURE PUBLISHING GROUP. ISSN 1750-2799

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Abstract

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. This protocol describes how to anesthetize and image different organs of Drosophila larvae, including epidermis, gut, imaginal discs, neurons, fat body, tracheae, muscles and hemocytes. It also explains how to use laser ablation to probe cell properties.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Kakanj, Parisa UNSPECIFIED UNSPECIFIED UNSPECIFIED Eming, Sabine A. UNSPECIFIED UNSPECIFIED UNSPECIFIED Partridge, Linda UNSPECIFIED orcid.org/0000-0001-9615-0094 UNSPECIFIED Leptin, Maria UNSPECIFIED orcid.org/0000-0001-7097-348X UNSPECIFIED
URN:	urn:nbn:de:hbz:38-345144
DOI:	10.1038/s41596-019-0282-z
Journal or Publication Title:	Nat. Protoc.
Volume:	15
Number:	3
Page Range:	S. 1158 - 1188
Date:	2020
Publisher:	NATURE PUBLISHING GROUP
Place of Publication:	LONDON
ISSN:	1750-2799
Language:	English
Faculty:	Faculty of Mathematics and Natural Sciences
Divisions:	Faculty of Mathematics and Natural Sciences > Department of Biology > Institute for Genetics
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language TARGETED GENE-EXPRESSION; CELL-SHAPE CHANGES; E-CADHERIN; MOSAIC ANALYSIS; TUMOR INVASION; ORGAN SIZE; PROTEIN; GROWTH; MODEL; JUNCTIONS Multiple languages Biochemical Research Methods Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/34514