利用确定性单细胞组合反应器高效组合多个单细胞。

IF 6.1 2区 工程技术 Q1 BIOCHEMICAL RESEARCH METHODS
Lab on a Chip Pub Date : 2024-12-16 DOI:10.1039/d4lc00951g
Mina Yoshida, Saori Tago, Kunihiko Iizuka, Teruo Fujii, Soo Hyeon Kim
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引用次数: 0

摘要

多个单细胞和/或单个微珠的分隔为单细胞转录组分析或细胞-细胞相互作用等高级生物研究带来了巨大的潜力。为确保分析的可靠性并防止误读,必须实现单个物体的高效配对或组合。在本文中,我们介绍了一种新型微流控装置,该装置与多层互联 Si/SiO2 控制电路相结合,被命名为确定性单细胞组合反应器(DSCR)装置,可实现多个单细胞的高效组合。多个单细胞的确定性组合是通过依次将每个细胞群引入并捕获到每个 DSCR 内指定的捕获孔中来实现的。这些细胞大小的陷阱孔是通过蚀刻二氧化硅钝化层形成的,可产生高度局部化的电场,从而促进确定性的单细胞捕获。该装置的多层电极互连实现了每个捕集阱的顺序操作,从而可以在组合反应器阵列中将每个细胞群精确捕集到指定的捕集阱中。我们通过依次诱捕三组不同的 PC3 细胞来证明 DSCR 的可行性,每组细胞都用不同的荧光染料(蓝色、绿色或红色)染色。这种方法在两个细胞群中的配对效率为 93 ± 2%,在三个细胞群中的组合效率为 82 ± 7%。我们的创新系统为分析多种细胞-细胞通信和单细胞组合索引提供了前景广阔的应用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Highly efficient combination of multiple single cells using a deterministic single-cell combinatorial reactor.

Compartmentalization of multiple single cells and/or single microbeads holds significant potential for advanced biological research including single-cell transcriptome analysis or cell-cell interactions. To ensure reliable analysis and prevent misinterpretation, it is essential to achieve highly efficient pairing or combining of single objects. In this paper, we introduce a novel microfluidic device coupled with a multilayer interconnect Si/SiO2 control circuit, named the deterministic single-cell combinatorial reactor (DSCR) device, for the highly efficient combination of multiple single cells. The deterministic combination of multiple single cells is realized by sequentially introducing and trapping each cell population into designated trap-wells within each DSCR. These cell-sized trap-wells, created by etching the SiO2 passivation layer, generate a highly localized electric field that facilitates deterministic single-cell trapping. The device's multilayer interconnection of electrodes enables the sequential operation of each trap-well, allowing precise trapping of each cell population into designated trap-wells within an array of combinatorial reactors. We demonstrated the feasibility of the DSCR by sequentially trapping three distinct groups of PC3 cells, each stained with a different fluorescent dye (blue, green, or red). This method achieved a 93 ± 2% pairing efficiency for two cell populations and an 82 ± 7% combination efficiency for three cell populations. Our innovative system offers promising applications for analyzing multiple cell-cell communications and combinatorial indexing of single cells.

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来源期刊
Lab on a Chip
Lab on a Chip 工程技术-化学综合
CiteScore
11.10
自引率
8.20%
发文量
434
审稿时长
2.6 months
期刊介绍: Lab on a Chip is the premiere journal that publishes cutting-edge research in the field of miniaturization. By their very nature, microfluidic/nanofluidic/miniaturized systems are at the intersection of disciplines, spanning fundamental research to high-end application, which is reflected by the broad readership of the journal. Lab on a Chip publishes two types of papers on original research: full-length research papers and communications. Papers should demonstrate innovations, which can come from technical advancements or applications addressing pressing needs in globally important areas. The journal also publishes Comments, Reviews, and Perspectives.
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