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气泡喷射式分拣器的耐用性使其能够实现高性能的生物样本分离。

Durability of the bubble-jet sorter enables high performance bio sample isolation.

机构信息

imec, Kapeldreef 75, 3001 Leuven, Belgium.

Department of Physics and Astronomy, KU Leuven, Celestijnenlaan 200D, 3001 Leuven, Belgium.

出版信息

Anal Methods. 2024 Sep 12;16(35):6030-6040. doi: 10.1039/d4ay01168f.

Abstract

Sorting cells while maintaining their viability for further processing or analysis is an essential step in a variety of biological processes ranging from early diagnostics to cell therapy. Sorting techniques such as fluorescence-activated cell sorting (FACS) have evolved considerably and provide standard ways of sorting. Nevertheless, the search for compact, integrated, efficient, and high throughput microfluidic sorting platforms continues due to challenges such as cost, cell viability, and biosafety. In our previous work, we introduced a technology with the potential to become such a platform: the bubble-jet sorter. It is a silicon-based sorter chip relying on cell deflection through micro vapor bubble formation. In this work, we present a new version of the sorter chip that emphasizes durability and continuous sorting operation. To characterize the sorter, we first focus on the technical performance and show a sorter lifetime that repeatedly exceeds 80 million actuation cycles. In addition, we show continuous operation at high firing rates, but also discuss limitations due to heat buildup. In a second step, we present continuous sorting runs of millions of beads and CD3 positive T cells at rates surpassing 1000 sorting events per second, while maintaining high purity (>90%) and recovery (>85%). Dedicated viability tests show that the gentle sorting process maintains cell viability in this closed, aerosol-free device. The remarkable combination of high lifetime, sorting rate, and sorting efficiency, along with the potential for on-chip parallelization show the promise of this technology to meet the growing demand for large-scale sample isolation in drug and immunotherapy development.

摘要

对细胞进行分选,同时保持其活力以进行进一步的处理或分析,是从早期诊断到细胞治疗等各种生物过程中的一个关键步骤。分选技术,如荧光激活细胞分选(FACS)已经有了很大的发展,并提供了标准的分选方法。然而,由于成本、细胞活力和生物安全性等挑战,人们仍在继续寻找紧凑、集成、高效和高通量的微流控分选平台。在我们之前的工作中,我们引入了一种有潜力成为这种平台的技术:气泡喷射式分选器。它是一种基于硅的分选芯片,依靠通过微蒸汽泡形成来使细胞发生偏转。在这项工作中,我们展示了一种新版本的分选芯片,该芯片强调耐用性和连续的分选操作。为了对分选器进行表征,我们首先关注技术性能,并展示了分选器寿命多次超过 8000 万次激活循环的结果。此外,我们还展示了在高触发率下的连续操作,但也讨论了由于热积累而导致的限制。在第二步中,我们展示了数以百万计的珠子和 CD3 阳性 T 细胞的连续分选运行,其分选速度超过每秒 1000 次分选事件,同时保持了>90%的高纯度和>85%的高回收率。专门的活力测试表明,在这个封闭的、无气溶胶的设备中,温和的分选过程保持了细胞活力。高寿命、分选速度和分选效率的显著结合,以及在芯片上进行并行化的潜力,表明这项技术有望满足药物和免疫疗法开发中对大规模样本分离的日益增长的需求。

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