Peninsula Medical School, University of Plymouth. John Bull Building, 16 Research Way, Plymouth PL6 8BU, UK.
Peninsula Medical School, University of Plymouth. John Bull Building, 16 Research Way, Plymouth PL6 8BU, UK.
STAR Protoc. 2022 Dec 16;3(4):101735. doi: 10.1016/j.xpro.2022.101735. Epub 2022 Sep 30.
Here, we describe a protocol to remove single identified cells directly from Drosophila living brains and analyze their transcriptome. We detail the steps to harvest fluorescent cells using a capillary under epifluorescence and transmitted light to avoid contamination. We then outline the procedure to obtain the transcriptome by reverse transcription and amplification. The process from cell harvesting to the initiation of reverse transcription only takes 2 min, thus avoiding transcriptional activation of cell damage response or cell death genes. For complete details on the use and execution of this protocol, please refer to Barros and Bossing (2021), Bossing et al. (2012), Gil-Ranedo et al. (2019), and Liu and Bossing (2016).
在这里，我们描述了一种从果蝇活体大脑中直接分离单个鉴定细胞并分析其转录组的方案。我们详细介绍了使用毛细管在明场和荧光下收获荧光细胞的步骤，以避免污染。然后，我们概述了通过反转录和扩增获得转录组的过程。从细胞收获到反转录开始的过程仅需 2 分钟，从而避免了细胞损伤反应或细胞死亡基因的转录激活。有关此方案使用和执行的完整详细信息，请参阅 Barros 和 Bossing（2021）、Bossing 等人（2012）、Gil-Ranedo 等人（2019）和 Liu 和 Bossing（2016）。
