Abstract：
Tissue dissociation, a crucial step in single-cell sample preparation, can alter the transcriptional state of a sample through the intrinsic cellular stress response. Here we demonstrate a general approach for measuring transcriptional response during sample preparation. In our method, transcripts made during dissociation are labeled for later identification upon sequencing. We found general as well as cell-type-specific dissociation response programs in zebrafish larvae, and we observed sample-to-sample variation in the dissociation response of mouse cardiomyocytes despite well-controlled experimental conditions. Finally, we showed that dissociation of the mouse hippocampus can lead to the artificial activation of microglia. In summary, our approach facilitates experimental optimization of dissociation procedures as well as computational removal of transcriptional perturbation response.
摘要：
组织解离，单细胞样品制备的关键步骤,可以通过内在的细胞应激反应改变样品的转录状态。在这里，我们展示了在样品制备过程中测量转录反应的一般方法。在我们的方法中，在解离过程中产生的转录物被标记用于以后测序时的鉴定。我们在斑马鱼幼虫中发现了一般以及细胞类型特异性的解离反应程序，尽管实验条件控制良好，但我们观察到小鼠心肌细胞解离反应的样品间变化。最后,我们发现小鼠海马的解离可以导致小胶质细胞的人工激活。总之,我们的方法促进了解离程序的实验优化以及转录扰动响应的计算去除。
