Department of Chemical and Biomolecular Engineering, University of Notre Dame, Notre Dame, IN, USA.
Bioengineering Graduate Program, University of Notre Dame, Notre Dame, IN, USA.
Methods Mol Biol. 2025;2861:257-271. doi: 10.1007/978-1-0716-4164-4_19.
Ex vivo calcium imaging in Drosophila opens an expansive amount of research avenues for the study of live signal propagation through complex tissue. Here, we describe how to isolate Drosophila organs of interest, like the developing wing imaginal disc and larval brain, culture them for extended periods, up to 10 h, and how to image the calcium dynamics occurring within them using genetically encoded biosensors like GCaMP. This protocol enables the study of complex calcium signaling dynamics, which is conserved throughout biology in such processes as cell differentiation and proliferation, immune reactions, wound healing, and cell-to-cell and organ-to-organ communication, among others. These methods also allow pharmacological compounds to be tested to observe effects on calcium dynamics with the applications of target identification and therapeutic development.
在果蝇中进行离体钙成像为研究活体信号通过复杂组织的传播开辟了广阔的研究途径。在这里，我们描述了如何分离果蝇感兴趣的器官，如发育中的翅膀 imaginal 盘和幼虫大脑，并对其进行长期培养，最长可达 10 小时，以及如何使用 GCaMP 等基因编码的生物传感器来成像它们内部发生的钙动力学。该方案可用于研究复杂的钙信号动力学，该动力学在生物学中在细胞分化和增殖、免疫反应、伤口愈合以及细胞间和器官间通讯等过程中是保守的。这些方法还允许测试药理学化合物，以观察对钙动力学的影响，从而进行靶标鉴定和治疗开发。
