Department of Biomedical Engineering, Erciyes University, 38039 Kayseri, Türkiye.
Nanothera Lab, Drug Application and Research Center (ERFARMA), Erciyes University, 38039 Kayseri, Türkiye.
Biosensors (Basel). 2022 Nov 13;12(11):1013. doi: 10.3390/bios12111013.
Simple staining of cells is a widely used method in basic medical diagnostics, education, and research laboratories. The stains are low-cost, but the extensive consumption results in excessive toxic waste generation. Thus, to decrease the amount of toxic waste resulting from the cell staining procedure is a need. In this study, we developed a magnetically driven and compartmentalized passive microfluidic chip to perform simple staining of human eukaryotic cells, K562 cells, and lymphocyte cells derived from patients. We demonstrated simple staining on cells with trypan blue, methylene blue, crystal violet, and safranin for high, medium, and low cell densities. The stained cells were imaged using a bright field optical microscope and a cell phone to count cells on the focal plane. The staining improved the color signal of the cell by 25-135-pixel intensity changes for the microscopic images. The validity of the protocol was determined using Jurkat and MDA-MB-231 cell lines as negative controls. In order to demonstrate the practicality of the system, lymphocyte cells derived from human blood samples were stained with trypan blue. The color intensity changes in the first and last compartments were analyzed to evaluate the performance of the chip. The developed method is ultra-low cost, significantly reduces the waste generated, and can be integrated with mobile imaging devices in terms of portability. By combining microfabrication technology with cell staining, this study reported a novel contribution to the field of microfluidic biosensors. In the future, we expect to demonstrate the detection of pathogens using this method.
细胞简单染色是基础医学诊断、教育和研究实验室中广泛使用的方法。这种染色剂成本低廉,但由于大量消耗,会产生过多的有毒废物。因此,减少细胞染色过程产生的有毒废物量是一种需求。在这项研究中,我们开发了一种基于磁场驱动和分隔的被动微流控芯片,用于对人类真核细胞、K562 细胞和源自患者的淋巴细胞进行简单染色。我们用台盼蓝、亚甲基蓝、结晶紫和番红对高、中、低细胞密度的细胞进行了简单染色。用明场光学显微镜和手机对染色细胞进行成像,以在焦平面上计数细胞。染色后,细胞的微观图像的颜色信号强度提高了 25-135 个像素。该方案的有效性通过 Jurkat 和 MDA-MB-231 细胞系作为阴性对照来确定。为了证明该系统的实用性,用人血样本中的淋巴细胞进行了台盼蓝染色。分析第一和最后一个隔室的颜色强度变化,以评估芯片的性能。所开发的方法成本极低,可显著减少产生的废物,并且在便携性方面可以与移动成像设备集成。通过将微制造技术与细胞染色相结合,本研究为微流控生物传感器领域做出了新的贡献。将来,我们期望使用这种方法来检测病原体。