A rapid, gentle and scalable method for dissociation and fluorescent sorting of imaginal disc cells for mRNA sequencing
A rapid, gentle and scalable method for dissociation and fluorescent sorting of imaginal disc cells for mRNA sequencing
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DOI：
10.1080/19336934.2016.1173296
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发表时间：
2016-01-01
期刊：
影响因子：
1.2
通讯作者：
Smith-Bolton, Rachel K.
中科院分区：
文献类型：
--
作者：
Khan, Sumbul Jawed;Abidi, Syeda Nayab Fatima;Smith-Bolton, Rachel K.
之前已经进行了成虫盘细胞的解离，以便能够通过流式细胞术和细胞分选来分析细胞周期进展、细胞大小、基因表达和成虫组织的其他方面。然而，所采用的冗长解离方案可能会改变基因表达、细胞行为和整体活力。在这里，我们描述了一种快速而温和的分离翼成虫盘细胞的方法，该方法可显着增强细胞活力并降低基因表达变化的可能性。此外，该方法具有可扩展性，能够收集大量样本进行高通量实验，而无需进行数据失真的扩增。
Dissociation of imaginal disc cells has been carried out previously to enable flow cytometry and cell sorting to analyze cell cycle progression, cell size, gene expression, and other aspects of imaginal tissues. However, the lengthy dissociation protocols employed may alter gene expression, cell behavior and overall viability. Here we describe a new rapid and gentle method of dissociating the cells of wing imaginal discs that significantly enhances cell viability and reduces the likelihood of gene expression changes. Furthermore, this method is scalable, enabling collection of large amounts of sample for high-throughput experiments without the need for data-distorting amplifications.