Azuaje-Hualde Enrique, Lartitegi Naiara, Benito-Lopez Fernando, Basabe-Desmonts Lourdes
Microfluidics Cluster UPV/EHU, BIOMICs Microfluidics Group, University of the Basque Country UPV/EHU, Vitoria-Gasteiz, Spain.
Microfluidics Cluster UPV/EHU, Analytical Microsystems & Materials for Lab-On-a-Chip (AMMa-LOAC) Group, University of the Basque Country UPV/EHU, Vitoria-Gasteiz, Spain.
Mikrochim Acta. 2025 Jul 5;192(8):484. doi: 10.1007/s00604-025-07319-2.
Accurately detecting cell secretions in complex microenvironments is crucial for understanding cellular communication, disease progression, and therapeutic responses. Traditional methods, such as ELISA, provide limited insight into the spatiotemporal dynamics of secretions, often requiring invasive or endpoint analyses. To address these challenges, we have functionalized the CellStudio platform-previously developed to integrate cell adhesion areas and microbeads patterns-with a novel self-reporting structure-switching signaling aptamer (SSSA) for vascular endothelial growth factor (VEGF) detection. This functionalization enables in situ single-step, real-time quantification of VEGF secretion from hundreds of live mesenchymal stem cell (MSC) clusters over a 24-h period, using standard fluorescence microscopy without the need for cell fixation. The spatial resolution provided by this system enables precise, localized monitoring of secretion events without the need to add extra reagents. This user-friendly platform is easily adaptable to conventional laboratory workflows, offering a versatile tool with potential for studying cellular behavior in real time with minimal technical barriers.
在复杂的微环境中准确检测细胞分泌物对于理解细胞通讯、疾病进展和治疗反应至关重要。传统方法，如酶联免疫吸附测定（ELISA），对分泌物的时空动态提供的见解有限，通常需要侵入性或终点分析。为应对这些挑战，我们对先前开发的用于整合细胞粘附区域和微珠模式的CellStudio平台进行了功能化，采用一种新型的自我报告结构转换信号适体（SSSA）来检测血管内皮生长因子（VEGF）。这种功能化能够在24小时内，使用标准荧光显微镜对数百个活的间充质干细胞（MSC）簇分泌的VEGF进行原位单步实时定量，而无需细胞固定。该系统提供的空间分辨率能够精确、局部地监测分泌事件，无需添加额外试剂。这个用户友好的平台很容易适应传统实验室工作流程，提供了一种通用工具，有可能以最小的技术障碍实时研究细胞行为。
