BIOS, Lab on a Chip group, MESA⁺ Institute for Nanotechnology, University of Twente, P.O. Box 217, 7500 AE Enschede, The Netherlands.
Analyst. 2013 Aug 21;138(16):4585-92. doi: 10.1039/c3an00569k. Epub 2013 Jun 10.
Today, droplet based microfluidics has become a standard platform for high-throughput single cell experimentation and analysis. However, until now no label-free, integrated single cell detection and discrimination method in droplets is available. We present here a microfluidic chip for fast (>100 Hz) and label-free electrical impedance based detection of cells in droplets. The microfluidic glass-PDMS device consists of two main components, the droplet generator and the impedance sensor. The planar electrode pair in the main channel allows the detection of only cells and cell containing droplets passing the electrodes using electrical impedance measurements. At a measurement frequency of 100 kHz non-viable cells, in low-conducting (LC) buffer, show an increase in impedance, due to the resistive effect of the membrane. The opposite effect, an impedance decrease, was observed when a viable cell passed the electrode pair, caused by the presence of the conducting cytoplasm. Moreover, we found that the presence of a viable cell in a droplet also decreased the measured electrical impedance. This impedance change was not visible when a droplet containing a non-viable cell or an empty droplet passed the electrode pair. A non-viable cell in a droplet and an empty droplet were equally classified. Hence, droplets containing (viable) cells can be discriminated from empty droplets. In conclusion, these results provide us with a valuable method to label-free detect and select viable cells in droplets. Furthermore, the proposed method provides the first step towards additional information regarding the encapsulated cells (e.g., size, number, morphology). Moreover, this all-electric approach allows for all-integrated Lab on a Chip (LOC) devices for cell applications using droplet-based platforms.
如今,液滴微流控已成为高通量单细胞实验和分析的标准平台。然而,到目前为止,还没有无标记的、集成的单细胞在液滴中的检测和区分方法。我们在这里提出了一种用于快速(>100 Hz)和无标记的基于电阻抗的液滴中细胞检测的微流控芯片。该微流控玻璃-聚二甲基硅氧烷(PDMS)器件由两个主要部分组成,即液滴发生器和阻抗传感器。主通道中的平面电极对仅允许通过电极的细胞和含有细胞的液滴进行细胞检测,这是通过测量电阻抗来实现的。在 100 kHz 的测量频率下,非存活细胞在低传导(LC)缓冲液中表现出阻抗增加,这是由于细胞膜的电阻效应。当一个存活细胞通过电极对时,观察到相反的效应,即阻抗降低,这是由于细胞质的存在导致的。此外,我们发现当一个含有存活细胞的液滴通过电极对时,测量的电阻抗也会降低。当一个含有非存活细胞的液滴或一个空液滴通过电极对时,这种阻抗变化是不可见的。因此,可以区分含有(存活)细胞的液滴和空液滴。总之,这些结果为我们提供了一种有价值的无标记方法,可以在液滴中检测和选择存活细胞。此外,该方法为进一步获取有关封装细胞的信息(例如,大小、数量、形态)提供了第一个步骤。此外,这种全电方法允许使用基于液滴的平台,为基于芯片的实验室(LOC)设备集成所有功能。