Advanced Optical Microscopy Unit, Central Core Facilities, Instituto de Salud Carlos III (ISCIII), Madrid, Spain.
Methods Mol Biol. 2025;2871:155-161. doi: 10.1007/978-1-0716-4217-7_14.
Microfluidics has emerged as a novel tool in the study of cellular dynamics, offering a precise control of cell microenvironment and real-time analysis of intracellular processes. This chapter explores the application of microfluidic technology in the translocation dynamics of the forkhead box O3 (FOXO3) transcription factor. FOXO3 is regulator of cellular homeostasis, apoptosis, and oxidative stress responses. By integrating microfluidic devices with live cell imaging techniques, we achieve fine-tuned manipulation of extracellular conditions, including pharmacological treatments, to observe their effects on FOXO3 localization at single-cell resolution. Microfluidic devices provide a versatile and precise platform for studying dynamic cellular processes such as FOXO3-GFP translocation. This methodology allows for the real-time observation and quantification of protein translocation in response to various stimuli, enhancing our understanding of cellular signaling mechanisms.
微流控技术已成为研究细胞动力学的新工具,可精确控制细胞微环境并实时分析细胞内过程。本章探讨了微流控技术在叉头框 O3(FOXO3)转录因子易位动力学中的应用。FOXO3 是细胞内稳态、细胞凋亡和氧化应激反应的调节剂。通过将微流控器件与活细胞成像技术相结合,我们可以精细地操控细胞外环境,包括药理学处理,以观察它们对 FOXO3 在单细胞分辨率下定位的影响。微流控器件为研究 FOXO3-GFP 易位等动态细胞过程提供了一个通用且精确的平台。这种方法可以实时观察和量化蛋白质易位对各种刺激的反应,增强我们对细胞信号转导机制的理解。