Microfluidics-free single-cell genomics with templated emulsification.
Microfluidics-free single-cell genomics with templated emulsification.
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DOI：
10.1038/s41587-023-01685-z
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发表时间：
2023-11
影响因子：
46.9
通讯作者：
Abate, Adam R.
中科院分区：
文献类型：
--
作者：
Clark, Iain C.;Fontanez, Kristina M.;Meltzer, Robert H.;Xue, Yi;Hayford, Corey;May-Zhang, Aaron;D'Amato, Chris;Osman, Ahmad;Zhang, Jesse Q.;Hettige, Pabodha;Ishibashi, Jacob S. A.;Delley, Cyrille L.;Weisgerber, Daniel W.;Replogle, Joseph M.;Jost, Marco;Phong, Kiet T.;Kennedy, Vanessa E.;Peretz, Cheryl A. C.;Kim, Esther A.;Song, Siyou;Karlon, William;Weissman, Jonathan S.;Smith, Catherine C.;Gartner, Zev J.;Abate, Adam R.
目前单细胞rna测序方法的局限性源于样品处理所需的微流体装置或流体处理步骤。我们开发了一种不需要专门的微流体设备，专业知识或硬件的方法。我们的方法是基于颗粒模板乳化，它允许单细胞封装和cDNA条形码在均匀的液滴乳剂只有一个漩涡。颗粒模板即时分割测序（PIP-seq）适用于多种乳化形式，包括微孔板和大体积锥形管，可在几分钟内处理数千个样品或数百万个细胞。我们证明PIP-seq在小鼠-人混合研究中产生高纯度的转录组，与多组学测量相兼容，与商业微流控平台相比，可以准确表征人类乳腺组织中的细胞类型。使用PIP-seq分析混合表型急性白血病的单细胞转录谱，揭示了标准免疫表型所隐藏的化疗耐药细胞亚群中异质性的出现。PIP-seq是一种简单，灵活和可扩展的下一代工作流程，可将单细胞测序扩展到新的应用。无微流体，可扩展的单细胞rna测序方法产生高质量的转录组。
Current single-cell RNA-sequencing approaches have limitations that stem from the microfluidic devices or fluid handling steps required for sample processing. We develop a method that does not require specialized microfluidic devices, expertise or hardware. Our approach is based on particle-templated emulsification, which allows single-cell encapsulation and barcoding of cDNA in uniform droplet emulsions with only a vortexer. Particle-templated instant partition sequencing (PIP-seq) accommodates a wide range of emulsification formats, including microwell plates and large-volume conical tubes, enabling thousands of samples or millions of cells to be processed in minutes. We demonstrate that PIP-seq produces high-purity transcriptomes in mouse–human mixing studies, is compatible with multiomics measurements and can accurately characterize cell types in human breast tissue compared to a commercial microfluidic platform. Single-cell transcriptional profiling of mixed phenotype acute leukemia using PIP-seq reveals the emergence of heterogeneity within chemotherapy-resistant cell subsets that were hidden by standard immunophenotyping. PIP-seq is a simple, flexible and scalable next-generation workflow that extends single-cell sequencing to new applications. A microfluidics-free, scalable single-cell RNA-sequencing method produces high-quality transcriptomes.
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微流体单细胞基因组学的模块化条形码珠。
影响因子：
4.6
作者：
Delley CL;Abate AR
通讯作者：
Abate AR
影响因子：
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作者：
Fancello L;Kampen KR;Hofman IJ;Verbeeck J;De Keersmaecker K
通讯作者：
De Keersmaecker K
影响因子：
64.8
作者：
Cao, Junyue;Spielmann, Malte;Shendure, Jay
通讯作者：
Shendure, Jay
影响因子：
6.1
作者：
Clark IC;Abate AR
通讯作者：
Abate AR
影响因子：
7.4
作者：
DeJournette, Cheryl J.;Kim, Joonyul;Medlen, Haley;Li, Xiangpeng;Vincent, Luke J.;Easley, Christopher J.
通讯作者：
Easley, Christopher J.