Experimental Considerations for Single-Cell RNA Sequencing Approaches.
Experimental Considerations for Single-Cell RNA Sequencing Approaches.
复制标题
DOI：
10.3389/fcell.2018.00108
复制
发表时间：
2018
影响因子：
5.5
通讯作者：
Kessenbrock K
中科院分区：
文献类型：
--
作者：
Nguyen QH;Pervolarakis N;Nee K;Kessenbrock K
单细胞转录组学技术已经成为以单个细胞的分辨率探索细胞异质性的强大工具。细胞生物学中的先前科学知识在很大程度上限于通过批量分析方法生成的数据，其仅提供通常掩盖细胞异质性的平均读数。当感兴趣的生物效应仅限于给定组织内的细胞亚群（例如干细胞/祖细胞）或浸润肿瘤的免疫细胞亚群时，这种平均方法特别成问题。单细胞RNA测序（scRNAseq）的巨大进步使科学家能够克服这一限制，并允许深入研究以前未探索的稀有细胞类型。由于scRNAseq的高灵敏度，必须对实验设置和执行给予足够的关注。仔细处理和加工scRNAseq细胞对于保留天然表达谱至关重要，这将确保有意义的分析和结论。在这里，我们描述了典型的单细胞分析工作流程的各个步骤，从组织采购，细胞制备，到平台选择和数据分析，我们讨论了每个步骤中的关键挑战，这将作为一个有用的指南来导航单细胞测序的复杂领域。
Single-cell transcriptomic technologies have emerged as powerful tools to explore cellular heterogeneity at the resolution of individual cells. Previous scientific knowledge in cell biology is largely limited to data generated by bulk profiling methods, which only provide averaged read-outs that generally mask cellular heterogeneity. This averaged approach is particularly problematic when the biological effect of interest is limited to only a subpopulation of cells such as stem/progenitor cells within a given tissue, or immune cell subsets infiltrating a tumor. Great advances in single-cell RNA sequencing (scRNAseq) enabled scientists to overcome this limitation and allow for in depth interrogation of previously unexplored rare cell types. Due to the high sensitivity of scRNAseq, adequate attention must be put into experimental setup and execution. Careful handling and processing of cells for scRNAseq is critical to preserve the native expression profile that will ensure meaningful analysis and conclusions. Here, we delineate the individual steps of a typical single-cell analysis workflow from tissue procurement, cell preparation, to platform selection and data analysis, and we discuss critical challenges in each of these steps, which will serve as a helpful guide to navigate the complex field of single-cell sequencing.
登录
查看更多内容
影响因子：
5.4
作者：
Alles J;Karaiskos N;Praktiknjo SD;Grosswendt S;Wahle P;Ruffault PL;Ayoub S;Schreyer L;Boltengagen A;Birchmeier C;Zinzen R;Kocks C;Rajewsky N
通讯作者：
Rajewsky N
影响因子：
14.8
作者：
Krishnaswami SR;Grindberg RV;Novotny M;Venepally P;Lacar B;Bhutani K;Linker SB;Pham S;Erwin JA;Miller JA;Hodge R;McCarthy JK;Kelder M;McCorrison J;Aevermann BD;Fuertes FD;Scheuermann RH;Lee J;Lein ES;Schork N;McConnell MJ;Gage FH;Lasken RS
通讯作者：
Lasken RS
影响因子：
8.8
作者：
Baldan, V.;Griffiths, R.;Hawkins, R. E.;Gilham, D. E.
通讯作者：
Gilham, D. E.
影响因子：
16.6
作者：
Gao R;Kim C;Sei E;Foukakis T;Crosetto N;Chan LK;Srinivasan M;Zhang H;Meric-Bernstam F;Navin N
通讯作者：
Navin N
单细胞染色质可及性揭示了调节变化的原理。
影响因子：
64.8
作者：
Buenrostro JD;Wu B;Litzenburger UM;Ruff D;Gonzales ML;Snyder MP;Chang HY;Greenleaf WJ
通讯作者：
Greenleaf WJ