Line-Focused Optical Excitation of Parallel Acoustic Focused Sample Streams for High Volumetric and Analytical Rate Flow Cytometry,Analytical Chemistry

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Line-Focused Optical Excitation of Parallel Acoustic Focused Sample Streams for High Volumetric and Analytical Rate Flow Cytometry
Analytical Chemistry ( IF 6.7 ) Pub Date : 2017-09-05 00:00:00 , DOI: 10.1021/acs.analchem.7b02319
Daniel M. Kalb ₁ , Frank A. Fencl ₁ , Travis A. Woods _{1,

2} , August Swanson ₃ , Gian C. Maestas ₁ , Jaime J. Juárez ₁ , Bruce S. Edwards ₂ , Andrew P. Shreve ₁ , Steven W. Graves ₁

Affiliation

Flow cytometry provides highly sensitive multiparameter analysis of cells and particles but has been largely limited to the use of a single focused sample stream. This limits the analytical rate to ∼50K particles/s and the volumetric rate to ∼250 μL/min. Despite the analytical prowess of flow cytometry, there are applications where these rates are insufficient, such as rare cell analysis in high cellular backgrounds (e.g., circulating tumor cells and fetal cells in maternal blood), detection of cells/particles in large dilute samples (e.g., water quality, urine analysis), or high-throughput screening applications. Here we report a highly parallel acoustic flow cytometer that uses an acoustic standing wave to focus particles into 16 parallel analysis points across a 2.3 mm wide optical flow cell. A line-focused laser and wide-field collection optics are used to excite and collect the fluorescence emission of these parallel streams onto a high-speed camera for analysis. With this instrument format and fluorescent microsphere standards, we obtain analysis rates of 100K/s and flow rates of 10 mL/min, while maintaining optical performance comparable to that of a commercial flow cytometer. The results with our initial prototype instrument demonstrate that the integration of key parallelizable components, including the line-focused laser, particle focusing using multinode acoustic standing waves, and a spatially arrayed detector, can increase analytical and volumetric throughputs by orders of magnitude in a compact, simple, and cost-effective platform. Such instruments will be of great value to applications in need of high-throughput yet sensitive flow cytometry analysis.

中文翻译：

线聚焦光学激发的平行声聚焦样品流，用于高体积和分析速率流式细胞术

流式细胞仪可对细胞和颗粒进行高灵敏度的多参数分析，但在很大程度上仅限于使用单个聚焦样品流。这将分析速率限制为〜50K颗粒/ s，并将体积速率限制为〜250μL/ min。尽管有流式细胞术的分析能力，但仍有一些应用这些速率不足的情况，例如高细胞背景下的稀有细胞分析（例如，母体血液中的循环肿瘤细胞和胎儿细胞），大稀释样品中细胞/颗粒的检测（例如水质，尿液分析）或高通量筛选应用。在这里，我们报告了一种高度平行的声波流式细胞仪，该仪使用声驻波将粒子聚焦到2.3毫米宽的光学流通池中的16个平行分析点中。使用线聚焦激光和广角收集光学器件来激发并收集这些平行流的荧光发射到高速相机上进行分析。通过这种仪器格式和荧光微球标准品，我们获得了100K / s的分析速率和10 mL / min的流速，同时保持了与商用流式细胞仪相当的光学性能。我们最初的原型仪器的结果表明，关键的可并行化组件（包括线聚焦激光器，使用多节点声驻波的颗粒聚焦以及空间阵列检测器）的集成可以在紧凑的条件下将分析和体积通量提高几个数量级。，简单且具有成本效益的平台。

更新日期：2017-09-06

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