IIT Bombay, Department of Chemical Engineering, IIT Bombay, Powai, Mumbai, 400076, Maharashtra, India.
Biosens Bioelectron. 2022 Sep 1;211:114334. doi: 10.1016/j.bios.2022.114334. Epub 2022 May 10.
Flow cytometry is an essential analytical technique used in biomedical diagnostics to measure properties of cells, micro-organisms, and particles. Laser light is scattered from particles focused in a flow cell and collected by light sensors, where the intensity of the scattered light is a function of the scattering angle, the refractive index of the particle and surrounding medium, the wavelength of light, and the size and the shape of the particle. One of the critical parts of the cytometer is the flow cell where the particle stream is constrained into a tight region within 10-30 μm using hydrodynamic focusing. The conventional flow cells use thick quartz flow cells, which are expensive and therefore not suitable for instruments targeted for resource-constrained settings. We demonstrate a compact, economical, bio-compatible flow cell assembly design that incorporates inexpensive and easily available capillaries attached to sturdy polymer fixtures in a simple manner that performs the focusing of a sample stream of particles. The flow cell has been tested by studying the relation between sample core diameter, and sample and sheath flow rates. Small-angle scattering (forward scatter) and wide-angle scattering (side scatter) have been captured for the enumeration and characterization of particles. We show excellent agreement between the size distribution obtained via direct imaging and that obtained from light scattering. The flow cell was also used to successfully size white blood cells in human blood samples.
流式细胞术是一种用于生物医学诊断的重要分析技术,用于测量细胞、微生物和颗粒的特性。激光光从聚焦在流动细胞中的颗粒散射,并被光传感器收集,其中散射光的强度是散射角、颗粒和周围介质的折射率、光的波长以及颗粒的大小和形状的函数。细胞仪的关键部件之一是流动池,其中使用流体力聚焦将颗粒流约束在 10-30 μm 的紧密区域内。传统的流动池使用厚的石英流动池,这种流动池昂贵,因此不适合针对资源有限环境的仪器。我们展示了一种紧凑、经济、生物兼容的流动池组件设计,该设计采用简单的方式将廉价且易于获得的毛细管固定在坚固的聚合物夹具上,从而实现了颗粒样品流的聚焦。已经通过研究样品芯直径以及样品和鞘液流速之间的关系来测试该流动池。已经捕获了小角度散射(前向散射)和大角度散射(侧向散射),以用于对颗粒进行计数和特性描述。我们展示了通过直接成像获得的尺寸分布与通过光散射获得的尺寸分布之间的极好一致性。该流动池还成功用于对人血样中的白细胞进行尺寸测量。