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.

中文翻译：

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果蝇幼虫的长期体内成像。

果蝇幼虫已被用于研究细胞生物学中的许多过程，包括形态发生，生理学以及对药物和新治疗化合物的反应。尽管它具有作为模型系统的巨大潜力，但是由于缺乏有效的方法来长期固定幼虫，因此长期的实时成像在技术上一直具有挑战性。我们在这里描述了一种简单的麻醉方法，可以对表皮和其他幼虫器官（包括肠，假体盘，神经元，脂肪体，气管，肌肉和血细胞）进行麻醉并进行不间断的长期体内成像，长达8小时。我们还包括通过激光消融探测细胞特性的程序。我们提供了对不同麻醉药作用的调查，证明了短期暴露于乙醚对于长期固定幼虫是最有效的。除了基本的果蝇遗传学和饲养以及共聚焦显微镜，该方案不需要专门知识。它可以对幼虫的许多系统和亚细胞过程进行高分辨率研究。