Cheon Hojin, Kumar Samir, Lee Inha, Shin Sanghoon, Jang Hyeji, Lee Young-Sun, Nam Myung-Hyun, Jun Hyun Sik, Seo Sungkyu
Department of Electronics and Information Engineering, Korea University, Sejong 30019, Republic of Korea.
Department of Biotechnology and Bioinformatics, Korea University, Sejong 30019, Republic of Korea.
Biosensors (Basel). 2025 Feb 24;15(3):143. doi: 10.3390/bios15030143.
Morphological changes in leukocytes are valuable markers for diseases and immune responses. In our earlier work, we presented Cellytics, a device that uses lens-free shadow imaging technology (LSIT) to monitor natural killer cell activity. Here, we present an improved Cellytics system that has been upgraded to a four-channel configuration to achieve higher throughput while maintaining robust reproducibility for rapid and cost-effective leukocyte analysis. The performance of this multi-channel Cellytics system was improved through refinements to the micro-pinhole chip. Etched pinholes provided better image resolution and clarity compared to drilled pinholes. To stimulate leukocytes, we used an activation stimulator cocktail (ASC) and quantified the resulting morphological changes using shadow-based metrics, including peak-to-peak distance (PPD) and maxima-to-minima standard deviation (MMD-SD). In addition, we developed a new leukocyte activation parameter (LAP) to specifically assess these activation-induced morphological changes. After ASC stimulation, leukocytes showed significantly increased PPD and LAP values and decreased MMD-SD compared to non-activated leukocytes. These results are consistent with the results of the flow cytometric analysis. These results emphasize the potential of Cellytics for the rapid and accurate assessment of leukocyte activation and provide a valuable tool for both clinical diagnostics and basic immunological research.
白细胞的形态变化是疾病和免疫反应的重要标志物。在我们早期的工作中，我们展示了Cellytics，一种使用无透镜阴影成像技术（LSIT）来监测自然杀伤细胞活性的设备。在此，我们展示了一种改进的Cellytics系统，该系统已升级为四通道配置，以实现更高的通量，同时在快速且经济高效的白细胞分析中保持强大的可重复性。通过对微针孔芯片的改进，提高了这种多通道Cellytics系统的性能。与钻孔针孔相比，蚀刻针孔提供了更好的图像分辨率和清晰度。为了刺激白细胞，我们使用了激活刺激剂混合物（ASC），并使用基于阴影的指标（包括峰峰值距离（PPD）和最大值到最小值标准差（MMD-SD））对由此产生的形态变化进行量化。此外，我们开发了一种新的白细胞激活参数（LAP）来专门评估这些激活诱导的形态变化。与未激活的白细胞相比，ASC刺激后，白细胞的PPD和LAP值显著增加，MMD-SD降低。这些结果与流式细胞术分析结果一致。这些结果强调了Cellytics在快速准确评估白细胞激活方面的潜力，并为临床诊断和基础免疫学研究提供了一个有价值的工具。
