Small volume low mechanical stress cytometry using computer-controlled Braille display microfluidics
Small volume low mechanical stress cytometry using computer-controlled Braille display microfluidics
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DOI：
10.1039/b708187a
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发表时间：
2007-01-01
期刊：
影响因子：
6.1
通讯作者：
Takayama, Shuichi
中科院分区：
文献类型：
--
作者：
Tung, Yi-Chung;Torisawa, Yu-Suke;Takayama, Shuichi
本文介绍了一种用于对含有少量珍贵细胞的样品进行高效、无损分析的微流式细胞仪系统。该系统利用盲文显示针的驱动进行微尺度流体操作，并利用带CCD相机的荧光显微镜进行光学检测。微流控芯片是完全一次性的，由具有微通道特征的聚二甲基硅氧烷（PDMS）板组成，密封在可变形的PDMS薄膜上。该通道设计了扩散器，以减轻基于销动器的蠕动泵浦方案固有的脉动流动行为，以最大限度地提高流体动力聚焦样品，并在通道内的层流中产生最小的干扰。连接微流控通道的漏斗是为了高效装载少量细胞的样品而设计的，并且也位于芯片上，以防止在驱动过程中盲文针的挤压作用对样品的物理损坏。通过计算流体动力学（CFD）模拟和实验观察对试样加载方案进行了表征。首先使用荧光素溶液进行流场研究，然后使用已知相对强度的荧光珠进行光学检测性能校准。利用小鼠成肌细胞（C2C12）来研究该装置上样方案的细胞活力。此外，用低渗DNA染色缓冲液染色的人早幼粒细胞白血病（HL60）细胞也在系统中进行了细胞周期分析。通过分析来自小鼠胚胎干细胞的单个胚状体的细胞，证明了有效分析细胞数量较少的细胞样本的能力。因此，本文所设计的微流控装置有望实现易于使用、小样本量的流式细胞分析，并有可能与其他基于盲文显示的微流控装置进一步集成，以促进多功能芯片实验室用于哺乳动物细胞操作。
This paper describes a micro flow cytometer system designed for efficient and non-damaging analysis of samples with small numbers of precious cells. The system utilizes actuation of Braille-display pins for micro-scale fluid manipulation and a fluorescence microscope with a CCD camera for optical detection. The microfluidic chip is fully disposable and is composed of a polydimethylsiloxane (PDMS) slab with microchannel features sealed against a thin deformable PDMS membrane. The channels are designed with diffusers to alleviate pulsatile flow behaviors inherent in pin actuator- based peristaltic pumping schemes to maximize hydrodynamic focusing of samples with minimal disturbances in the laminar streams within the channel. A funnel connected to the microfluidic channel is designed for efficient loading of samples with small number of cells and is also positioned on the chip to prevent physical damages of the samples by the squeezing actions of Braille pins during actuation. The sample loading scheme was characterized by both computational fluidic dynamics (CFD) simulation and experimental observation. A fluorescein solution was first used for flow field investigation, followed by use of fluorescence beads with known relative intensities for optical detection performance calibration. Murine myoblast cells (C2C12) were exploited to investigate cell viability for the sample loading scheme of the device. Furthermore, human promyelocytic leukemia (HL60) cells stained by hypotonic DNA staining buffer were also tested in the system for cell cycle analysis. The ability to efficiently analyze cellular samples where the number of cells is small was demonstrated by analyzing cells from a single embryoid body derived from mouse embryonic stem cells. Consequently, the designed microfluidic device reported in this paper is promising for easy-to-use, small sample size flow cytometric analysis, and has potential to be further integrated with other Braille display-based microfluidic devices to facilitate a multi-functional lab-on-a-chip for mammalian cell manipulations.