Using Mosaic Cell Labeling to Visualize Polyploid Cells in the Drosophila Brain.
Using Mosaic Cell Labeling to Visualize Polyploid Cells in the Drosophila Brain.
复制标题
使用镶嵌细胞标记可视化果蝇大脑中的多倍体细胞。
DOI：
10.1007/978-1-0716-2561-3_22
复制
发表时间：
2023
期刊：
影响因子：
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通讯作者：
Buttitta,Laura
中科院分区：
文献类型：
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作者：
Nandakumar,Shyama;Buttitta,Laura
用于研究内复制的传统方法在用于识别复杂、密集组织中罕见的多倍化事件时存在局限性。在这里，我们描述了一种使用名为“CoinFLP”的现有镶嵌多色标记技术原位识别和可视化多倍体细胞的方法（Bosch et al., Development 142(3):597–606, 2015）。 CoinFLP 可以轻松地对多倍体细胞进行原位可视化，并且可以与其他技术相结合，例如用于细胞类型特异性标记的免疫荧光和流式细胞术来执行定量，也可用于遗传操作。此外，通过修改标记时间，该技术还可用于区分细胞融合事件和内周期事件（Nandakumar 等人，eLife 25:9, 2020），从而可以推断多倍化的方法。
Traditional methods used to study endoreplication have limitations when used to identify rare events of polyploidization in complex, densely-packed tissues. Here, we describe a method to identify and visualize polyploid cells in situ using an existing mosaic, multicolor labeling technique named “CoinFLP” (Bosch et al., Development 142(3):597–606, 2015). CoinFLP allows easy visualization of polyploid cellsin situand can be combined with other techniques such as immunofluorescence for cell-type-specific labeling and flow cytometry to perform quantifications and can also be used for genetic manipulations. Further, by modifying the time of labeling, this technique can also be used to distinguish events of cell fusion from endocycle (Nandakumar et al., eLife 25:9, 2020)—allowing one to infer the method of polyploidization.