用于实时计数和高精度检索极少量细胞的微流体平台

IF 2.3 4区 工程技术 Q2 INSTRUMENTS & INSTRUMENTATION Microfluidics and Nanofluidics Pub Date : 2024-09-27 DOI:10.1007/s10404-024-02767-3
Buket Şahin, Begüm Şen Doğan, Ebru Özgür, Özge Zorlu, Ender Yıldırım, Haluk Külah
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引用次数: 0

摘要

近年来,在细胞生物学研究、诊断和个性化医疗中研究稀有细胞的需求推动下,单细胞分离和分析变得至关重要。然而,用于分离少量细胞的现有平台价格昂贵、劳动密集型,而且不能广泛使用。为了解决这个问题,我们提出了一种低成本、可重复的微流体平台,能够以高精度和最小的样品损失提取 1-100 个细胞。该系统利用二维流体动力聚焦芯片和移液器吸头作为细胞储液器,并通过柔性液压储液器(FHR)防止样本丢失。利用注射泵驱动的流动收集细胞,在显微镜下进行实时监测,并利用图像处理软件进行计数。为了验证该平台,MCF7 乳腺癌细胞通过了微通道,目标回收数量为 1 到 100 个细胞。1 个、10 个、50 个和 100 个目标细胞的平均检索细胞数分别为 1.0 ± 0.0、9.2 ± 2.4、46.0 ± 5.9 和 98.5 ± 6.2。对于 1、10、50 和 100 个目标细胞,代码计数与检索细胞数之间的平均偏差分别为 0 ± 0.6、-0.3 ± 1.7、-1.6 ± 0.9 和 3.9 ± 4.8,这证明了代码计数的准确性。整个过程不到 10 分钟,细胞计数与目标非常吻合,显示出很高的准确性。重要的是,处理后细胞活力不受影响。这种方法为精确的细胞计数和检索提供了一种经济、可靠的解决方案,适用于各种下游应用。
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A microfluidic platform for real-time enumeration and high accuracy retrieval of a very low number of cells

In recent years, single cell isolation and analysis have become crucial, driven by the need to study rare cells in cell biology research, diagnostics, and personalized medicine. However, existing platforms for isolating small cell numbers are expensive, labor-intensive, and not widely accessible. To address this, we present a low-cost, repeatable microfluidic platform capable of retrieving 1-100 cells with high accuracy and minimal sample loss. The system utilizes a 2D hydrodynamic focusing chip and a pipette tip as a cell reservoir, enhanced by a flexible hydraulic reservoir (FHR) to prevent sample loss. Cells are collected using a syringe pump-driven flow, monitored in real-time under a microscope, and counted using image processing software. To validate the platform, MCF7 breast cancer cells were passed through the microchannel, with target retrieval numbers ranging from 1 to 100 cells. The average retrieved cell count was found to be 1.0 ± 0.0, 9.2 ± 2.4, 46.0 ± 5.9 and 98.5 ± 6.2 for 1, 10, 50, and 100 targeted number of cells, respectively. The counting accuracy of the code was demonstrated by the average deviation between the code count and retrieved number of cells being 0 ± 0.6, -0.3 ± 1.7, -1.6 ± 0.9, and 3.9 ± 4.8, respectively for 1, 10, 50, and 100 targeted cells. The process took less than 10 min, with cell counts matching targets closely and demonstrating high accuracy. Importantly, cell viability remained unaffected post-process. This method offers a cost-effective, robust solution for precise cell counting and retrieval, suitable for various downstream applications.

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来源期刊
Microfluidics and Nanofluidics
Microfluidics and Nanofluidics 工程技术-纳米科技
CiteScore
4.80
自引率
3.60%
发文量
97
审稿时长
2 months
期刊介绍: Microfluidics and Nanofluidics is an international peer-reviewed journal that aims to publish papers in all aspects of microfluidics, nanofluidics and lab-on-a-chip science and technology. The objectives of the journal are to (1) provide an overview of the current state of the research and development in microfluidics, nanofluidics and lab-on-a-chip devices, (2) improve the fundamental understanding of microfluidic and nanofluidic phenomena, and (3) discuss applications of microfluidics, nanofluidics and lab-on-a-chip devices. Topics covered in this journal include: 1.000 Fundamental principles of micro- and nanoscale phenomena like, flow, mass transport and reactions 3.000 Theoretical models and numerical simulation with experimental and/or analytical proof 4.000 Novel measurement & characterization technologies 5.000 Devices (actuators and sensors) 6.000 New unit-operations for dedicated microfluidic platforms 7.000 Lab-on-a-Chip applications 8.000 Microfabrication technologies and materials Please note, Microfluidics and Nanofluidics does not publish manuscripts studying pure microscale heat transfer since there are many journals that cover this field of research (Journal of Heat Transfer, Journal of Heat and Mass Transfer, Journal of Heat and Fluid Flow, etc.).
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