影像盘
生物
果蝇属(亚属)
表皮(动物学)
幼虫
发育生物学
体内
细胞生物学
黑腹果蝇
形态发生
临床前影像学
烧蚀
解剖
医学
遗传学
植物
内科学
基因
作者
Parisa Kakanj,Sabine A. Eming,Linda Partridge,Maria Leptin
出处
期刊:Nature Protocols
[Springer Nature]
日期:2020-02-10
卷期号:15 (3): 1158-1187
被引量:31
标识
DOI:10.1038/s41596-019-0282-z
摘要
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.
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