Single-Cell Digital Lysates Generated by Phase-Switch Microfluidic Device Reveal Transcriptome Perturbation of Cell Cycle

	Single-Cell Digital Lysates Generated by Phase-Switch Microfluidic Device Reveal Transcriptome Perturbation of Cell Cycle
	Chen, Gina Y.; Chen, Yan; Millstein, Joshua; Liu, Yao; Chen, Xuelian; Stucky, Andres; Qu, Cunye; Fan, Jian-Bing; Chang, Xiao; Soleimany, Ava
刊名	ACS NANO
	2018
文献子类	期刊论文
英文摘要	With conventional gene expression profiling, information concerning cellular heterogeneity is often lost in the physical mixing and averaging of millions of cells. Single cell transcriptome analysis has the potential to address these issues. However, there is a need to determine how many cells are needed to draw meaningful conclusions in each single-cell study. Here, we introduce the concept of "digital lysate" for assessing cellular heterogeneity with a phase-switch microfluidic platform and apply it to construct a molecular map of transcriptome perturbation during the cell cycle. Using a phase-switch droplet microfluidic platform and next-generation sequencing, we obtained transcriptomes of single cells by random sampling. Digital lysates were generated by permutating and averaging multiple single-cell transcriptomes. In our studied cell populations, digital lysates converged to physical lysates (r = 0.93), and the sample-to-sample repeatability was comparable to that of conventional analysis of a physical lysate (r = 0.98). After determining the number of cells needed, single-cell transcriptomes were used to organize cells into a map by molecular similarity, and the map was validated by cell cycle-specific markers (p = 0.003). Cell cycle regulatory genes were inferred using this molecular map and verified with siRNA assays. The study described here provides an effective approach, the generation and analysis of digital lysates, to investigate cellular heterogeneity.
URL标识	查看原文
语种	英语
内容类型	期刊论文
源URL	[http://ir.siat.ac.cn:8080/handle/172644/14287]
专题	深圳先进技术研究院_医工所
推荐引用方式 GB/T 7714	Chen, Gina Y.,Chen, Yan,Millstein, Joshua,et al. Single-Cell Digital Lysates Generated by Phase-Switch Microfluidic Device Reveal Transcriptome Perturbation of Cell Cycle[J]. ACS NANO,2018.
APA	Chen, Gina Y..,Chen, Yan.,Millstein, Joshua.,Liu, Yao.,Chen, Xuelian.,...&Zhong, Jiang F..(2018).Single-Cell Digital Lysates Generated by Phase-Switch Microfluidic Device Reveal Transcriptome Perturbation of Cell Cycle.ACS NANO.
MLA	Chen, Gina Y.,et al."Single-Cell Digital Lysates Generated by Phase-Switch Microfluidic Device Reveal Transcriptome Perturbation of Cell Cycle".ACS NANO (2018).