μCB-seq: microfluidic cell barcoding and sequencing for high-resolution imaging and sequencing of single cells.
μCB-seq: microfluidic cell barcoding and sequencing for high-resolution imaging and sequencing of single cells.
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DOI：
10.1039/d0lc00169d
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发表时间：
2020-11-07
期刊：
影响因子：
6.1
通讯作者：
Streets A
中科院分区：
文献类型：
--
作者：
Chen TN;Gupta A;Zalavadia MD;Streets A
单细胞 RNA 测序 (scRNA-seq) 能够以高分辨率研究多细胞生物中的复杂生物过程。然而，对于理解异质组织或器官中细胞的功能作用至关重要的许多表型特征并未直接在基因组中编码，因此无法使用 scRNA-seq 进行分析。定量光学显微镜长期以来一直是表征不同细胞表型（包括细胞形态、蛋白质定位和化学成分）的有效方法。将 scRNA-seq 与光学成像相结合有可能提供全面的单细胞分析，从而实现基因表达谱和细胞状态表征的功能整合。然而，通过这两种测量来追踪单个细胞是很困难的；因此，将当前的 scRNA-seq 协议与光学测量结合起来仍然是一个挑战。在这里，我们报告了微流控细胞条形码和测序（μCB-seq），这是一种结合了单细胞高分辨率成像和测序的微流控平台。 μCB-seq 通过一种新颖的制造方法实现，该方法将具有已知条形码序列的引物预加载到微流体装置的可寻址反应室内。除了实现多模式单细胞分析之外，μCB-seq 还提高了基因检测灵敏度，为单细胞的信息丰富的表征提供了一种可扩展且准确的方法。
Single-cell RNA sequencing (scRNA-seq) enables the investigation of complex biological processes in multicellular organisms with high resolution. However, many phenotypic features that are critical to understanding the functional role of cells in a heterogeneous tissue or organ are not directly encoded in the genome and therefore cannot be profiled with scRNA-seq. Quantitative optical microscopy has long been a powerful approach for characterizing diverse cellular phenotypes including cell morphology, protein localization, and chemical composition. Combining scRNA-seq with optical imaging has the potential to provide comprehensive single-cell analysis, allowing for functional integration of gene expression profiling and cell-state characterization. However, it is difficult to track single cells through both measurements; therefore, coupling current scRNA-seq protocols with optical measurements remains a challenge. Here, we report Microfluidic Cell Barcoding and Sequencing (μCB-seq), a microfluidic platform that combines high-resolution imaging and sequencing of single cells. μCB-seq is enabled by a novel fabrication method that preloads primers with known barcode sequences inside addressable reaction chambers of a microfluidic device. In addition to enabling multi-modal single-cell analysis, μCB-seq improves gene detection sensitivity, providing a scalable and accurate method for information-rich characterization of single cells.
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影响因子：
5.8
作者：
Gu, Zuguang;Eils, Roland;Schlesner, Matthias
通讯作者：
Schlesner, Matthias
影响因子：
3.7
作者：
Bystrykh LV
通讯作者：
Bystrykh LV
DOI：
10.1126/science.1247651
发表时间：
2014-02-14
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Jaitin DA;Kenigsberg E;Keren-Shaul H;Elefant N;Paul F;Zaretsky I;Mildner A;Cohen N;Jung S;Tanay A;Amit I
通讯作者：
Amit I
影响因子：
46.9
作者：
通讯作者：
--
DOI：
10.1073/pnas.1507125112
发表时间：
2015-06-09
影响因子：
11.1
作者：
Darmanis S;Sloan SA;Zhang Y;Enge M;Caneda C;Shuer LM;Hayden Gephart MG;Barres BA;Quake SR
通讯作者：
Quake SR