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Serial flow cytometry in an inertial focusing optofluidic microchip for direct assessment of measurement variations
Lab on a Chip ( IF 6.1 ) Pub Date : 2022-07-20 , DOI: 10.1039/d1lc01169c Matthew DiSalvo 1, 2 , Paul N Patrone 3 , Anthony J Kearsley 3 , Gregory A Cooksey 2
Lab on a Chip ( IF 6.1 ) Pub Date : 2022-07-20 , DOI: 10.1039/d1lc01169c Matthew DiSalvo 1, 2 , Paul N Patrone 3 , Anthony J Kearsley 3 , Gregory A Cooksey 2
Affiliation
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Flow cytometry is an invaluable technology in biomedical research, but confidence in single-cell measurements remains limited due to a lack of appropriate techniques for uncertainty quantification (UQ). It is particularly challenging to evaluate the potential for different instrumentation designs or operating parameters to influence the measurement physics in ways that change measurement repeatability. Here, we report a direct experimental approach to UQ using a serial flow cytometer that measured each particle more than once along a flow path. The instrument was automated for real-time characterization of measurement precision and operated with particle velocities exceeding 1 m s−1, throughputs above 100 s−1, and analysis yields better than 99.9%. These achievements were enabled by a novel hybrid inertial and hydrodynamic particle focuser to tightly control particle positions and velocities. The cytometer identified ideal flow conditions with fluorescence area measurement precision on the order of 1% and characterized tradeoffs between precision, throughput, and analysis yield. The serial cytometer is anticipated to improve single-cell measurements through estimation (and subsequent control) of uncertainty contributions from various other instrument parameters leading to overall improvements in the ability to better classify sample composition and to find rare events.
中文翻译：
用于直接评估测量变化的惯性聚焦光流体微芯片中的串行流式细胞术
流式细胞术是生物医学研究中的一项宝贵技术，但由于缺乏适当的不确定性量化 (UQ) 技术，对单细胞测量的信心仍然有限。评估不同仪器设计或操作参数以改变测量重复性的方式影响测量物理的潜力尤其具有挑战性。在这里，我们报告了使用串行流式细胞仪对 UQ 进行直接实验的方法，该方法沿流路多次测量每个粒子。该仪器是自动化的，用于测量精度的实时表征，并以超过 1 ms -1的粒子速度运行，吞吐量超过 100 s -1, 分析结果优于 99.9%。这些成就是通过一种新型混合惯性和流体动力学粒子聚焦器来严格控制粒子位置和速度的。细胞仪确定了理想的流动条件，荧光面积测量精度约为 1%，并表征了精度、通量和分析产量之间的权衡。预计串行细胞仪将通过估计（和随后的控制）各种其他仪器参数的不确定性贡献来改进单细胞测量，从而全面提高更好地分类样品组成和发现罕见事件的能力。
更新日期：2022-07-20
中文翻译：
用于直接评估测量变化的惯性聚焦光流体微芯片中的串行流式细胞术
流式细胞术是生物医学研究中的一项宝贵技术，但由于缺乏适当的不确定性量化 (UQ) 技术，对单细胞测量的信心仍然有限。评估不同仪器设计或操作参数以改变测量重复性的方式影响测量物理的潜力尤其具有挑战性。在这里，我们报告了使用串行流式细胞仪对 UQ 进行直接实验的方法，该方法沿流路多次测量每个粒子。该仪器是自动化的，用于测量精度的实时表征，并以超过 1 ms -1的粒子速度运行，吞吐量超过 100 s -1, 分析结果优于 99.9%。这些成就是通过一种新型混合惯性和流体动力学粒子聚焦器来严格控制粒子位置和速度的。细胞仪确定了理想的流动条件，荧光面积测量精度约为 1%，并表征了精度、通量和分析产量之间的权衡。预计串行细胞仪将通过估计（和随后的控制）各种其他仪器参数的不确定性贡献来改进单细胞测量，从而全面提高更好地分类样品组成和发现罕见事件的能力。
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