A live-imaging protocol for tracking intestinal stem cell divisions in the Drosophila melanogaster pupal midgut.
A live-imaging protocol for tracking intestinal stem cell divisions in the Drosophila melanogaster pupal midgut.
复制标题
一个实时成像协议跟踪肠道干细胞分裂的果蝇蛹中肠。
DOI：
10.1016/j.xpro.2023.102749
复制
发表时间：
2023-12-15
期刊：
影响因子：
--
通讯作者：
Guo Z
中科院分区：
文献类型：
--
作者：
Wu S;Tang R;Ohlstein B;Guo Z
建立肠道干细胞（ISC）的长期离体观察对于帮助了解肠道上皮的形成和动态平衡至关重要。在这里，我们提出了一个协议，跟踪果蝇蛹ISCs蛹中肠发育过程中的分裂。我们描述的步骤解剖，安装，和现场成像蛹中肠。然后，我们详细说明每个细胞的荧光定量程序。该方案可以应用于其他荧光标记的蛋白质。有关本方案使用和执行的完整详细信息，请参见Wu等人。跟踪果蝇蛹肠细胞分裂的方案解剖蛹以获得蛹中肠蛹中肠的安装和蛹中肠的实时成像以及蛋白质定量出版商说明：进行任何实验方案都需要遵守当地机构的实验室安全和道德准则。建立肠干细胞（ISC）的长期离体观察对于帮助理解肠上皮的形成和稳态至关重要。在这里，我们提出了一个协议，跟踪果蝇蛹ISCs蛹中肠发育过程中的分裂。我们描述的步骤解剖，安装，和现场成像蛹中肠。然后，我们详细说明每个细胞的荧光定量程序。该方案可以应用于其他荧光标记的蛋白质。
Establishing a long-term ex vivo observation of the intestinal stem cell (ISC) is crucial to help understand the formation and homeostasis of the intestinal epithelium. Here, we present a protocol for tracking the division of Drosophila pupal ISCs during pupal midgut development. We describe steps for dissecting, mounting, and live imaging the pupal midgut. We then detail procedures for fluorescence quantification of each cell. This protocol can be applied to other fluorescently tagged proteins. For complete details on the use and execution of this protocol, please refer to Wu et al. Protocol for tracking the division of Drosophila pupal intestinal cells Dissection of the pupa to obtain the pupal midgut Mounting and live imaging of the pupal midgut and protein quantification Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Establishing a long-term ex vivo observation of the intestinal stem cell (ISC) is crucial to help understand the formation and homeostasis of the intestinal epithelium. Here, we present a protocol for tracking the division of Drosophila pupal ISCs during pupal midgut development. We describe steps for dissecting, mounting, and live imaging the pupal midgut. We then detail procedures for fluorescence quantification of each cell. This protocol can be applied to other fluorescently tagged proteins.