小柯机器人

美国加州大学旧金山分校Adam R. Abate团队近期取得重要工作进展，他们最新研究提出了使用具有模板乳化的无微流体单细胞基因组学方法。相关论文2023年3月6日在线发表于《自然—生物技术》杂志上。

据了解，当前的单细胞RNA测序方法具有来自微流体装置或样品处理所需的流体处理步骤的限制。

研究人员开发了一种新的方法，该方法不需要专门的微流体设备、专业知识或硬件。这种方法是基于粒子模板乳化，允许单细胞封装和条形码cDNA在均匀的液滴乳液中，只需涡旋器。颗粒模板化即时分区测序（PIP-seq）适应多种乳化形式，包括微孔板和大体积锥形管，使数千个样品或数百万个细胞在几分钟内得到处理。研究人员证明，PIP-seq在小鼠-人类混合研究中产生高纯度转录组，与多组学测量兼容，与商业微流体平台相比，可以准确地表征人类乳腺组织中的细胞类型。使用PIP-seq对混合表型急性白血病的单细胞转录谱分析揭示了标准免疫分型所隐藏的化疗耐药细胞亚群的异质性。

总之，PIP-seq是一个简单、灵活和可扩展的下一代工作流程，将单细胞测序扩展到新的应用程序。

附：英文原文

Title: Microfluidics-free single-cell genomics with templated emulsification

Author: Clark, Iain C., Fontanez, Kristina M., Meltzer, Robert H., Xue, Yi, Hayford, Corey, May-Zhang, Aaron, DAmato, 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.

Issue&Volume: 2023-03-06

Abstract: 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.

DOI: 10.1038/s41587-023-01685-z

Source: https://www.nature.com/articles/s41587-023-01685-z

Nature Biotechnology：《自然—生物技术》，创刊于1996年。隶属于施普林格·自然出版集团，最新IF：68.164
官方网址：https://www.nature.com/nbt/
投稿链接：https://mts-nbt.nature.com/cgi-bin/main.plex