Trauma and Transfusion Medicine Research Center, Department of Surgery, University of Pittsburgh, Pittsburgh, Pennsylvania.
Department of Chemical and Biological Engineering, The University of New Mexico, Albuquerque, New Mexico.
Curr Protoc. 2024 Oct;4(10):e1124. doi: 10.1002/cpz1.1124.
Flow cytometry is an inherently fluidic process that flows particles on a one-by-one basis through a sensing region to discretely measure their optical and physical properties. It can be used to analyze particles ranging in size from nanoparticles to whole organisms (e.g., zebrafish). It has particular value for blood analysis, and thus most instruments are fluidically optimized for particles that are comparable in size to a typical blood cell. The principles of fluid dynamics allow for particles of such size to be precisely positioned in flow as they pass through sensing regions that are tens of microns in length at linear velocities of meters per second. Such fluidic systems enable discrete analysis of cell-sized particles at rates approaching 100 kHz. For larger particles, the principles of fluidics greatly reduce the achievable rates, but such high rates of data acquisition for cell-sized particles allow rapid collection of information on many thousands to millions of cells and provides for research and clinical measurements of both rare and common cell populations with a high degree of statistical confidence. Additionally, flow cytometers can accurately count particles via the use of volumetric sample delivery and can be coupled with high-throughput sampling technologies to greatly increase the rate at which independent samples can be delivered to the system. Due to the combination of high analysis rates, sensitive multiparameter measurements, high-throughput sampling, and accurate counting, flow cytometry analysis is the gold standard for many critical applications in clinical, research, pharmaceutical, and environmental areas. Beyond the power of flow cytometry as an analytical technique, the fluidic pathway can be coupled with a sorting mechanism to collect particles based on desired properties. We present an overview of fluidic systems that enable flow cytometry-based analysis and sorting. We introduce historical approaches, explanations of commonly implemented fluidics, and brief discussions of potential future fluidics where appropriate. © 2024 Wiley Periodicals LLC.
流式细胞术是一种基于流体的过程,它以逐个粒子的方式将粒子流过感应区,以离散地测量其光学和物理特性。它可用于分析大小从纳米颗粒到整个生物体(例如斑马鱼)的粒子。它在血液分析方面具有特殊的价值,因此大多数仪器在流体方面都针对大小与典型血细胞相当的粒子进行了优化。流体动力学的原理允许在粒子以每秒米的线性速度通过长度为数十微米的感应区时,精确地将其定位在流中。这种流体系统能够以接近 100 kHz 的速率对细胞大小的粒子进行离散分析。对于较大的粒子,流体力学的原理大大降低了可实现的速率,但对于细胞大小的粒子如此高的数据采集速率允许快速收集数千到数百万个细胞的大量信息,并为稀有和常见细胞群体的研究和临床测量提供高度统计置信度。此外,流式细胞仪可以通过使用体积样品输送来准确地计数粒子,并可以与高通量采样技术结合使用,以大大提高可以输送到系统的独立样品的速率。由于高分析速率、敏感的多参数测量、高通量采样和准确计数的结合,流式细胞术分析是临床、研究、制药和环境领域许多关键应用的黄金标准。除了流式细胞术作为分析技术的强大功能外,流体路径还可以与分选机制结合使用,根据所需特性收集粒子。我们介绍了用于流式细胞术分析和分选的流体系统概述。我们介绍了历史方法、对常用流体学的解释以及在适当的情况下对潜在未来流体学的简要讨论。