Department of Anatomy, School of Medicine, University of California, San Francisco, CA, USA.
Methods Mol Biol. 2023;2626:323-333. doi: 10.1007/978-1-0716-2970-3_17.
The production of eggs in the Drosophila ovary requires complex interactions between multiple cell types that coexist within the same solid tissue. This cellular heterogeneity makes the ovary a rich subject of study, but also makes it challenging to identify transcriptional differences between individual cell types using methods such as bulk RNA sequencing. The development of single-cell RNA sequencing (scRNA-seq) techniques addresses this limitation by providing an avenue to profile genetic and functional heterogeneity at a cellular resolution. Here, we describe the isolation and preparation of the Drosophila ovary for scRNA-seq. This protocol emphasizes a short preparation time, high cell viability, prevention of RNA-degradation, and reduction of technical variation to achieve highly reproducible single-cell profiles.
果蝇卵巢中的卵子产生需要多种细胞类型之间的复杂相互作用，这些细胞类型共同存在于同一固体组织中。这种细胞异质性使卵巢成为一个丰富的研究课题，但也使得使用批量 RNA 测序等方法识别单个细胞类型之间的转录差异具有挑战性。单细胞 RNA 测序 (scRNA-seq) 技术的发展通过提供在细胞分辨率下对遗传和功能异质性进行分析的途径解决了这一限制。在这里，我们描述了果蝇卵巢用于 scRNA-seq 的分离和准备。该方案强调了短的准备时间、高细胞活力、防止 RNA 降解以及减少技术变异，以实现高度可重复的单细胞图谱。
