Instituto de Investigação e Inovação em Saúde and Instituto de Biologia Molecular e Celular, Universidade do Porto, 4200-135 Porto, Portugal.
Unit of Experimental Biology, Department of Biomedicine, Faculty of Medicine, University of Porto, 4200-319 Porto, Portugal.
STAR Protoc. 2020 Oct 23;1(3):100147. doi: 10.1016/j.xpro.2020.100147. eCollection 2020 Dec 18.
This protocol highlights the use of FRET-based biosensors to investigate signaling events during microglia activation in real time. Understanding microglia activation has gained momentum as it can help decipher signaling mechanisms underlying the neurodegenerative process occurring in neurological disorders. Unlike more traditional methods widely employed in the microglia field, FRET allows microglia signaling events to be studied in real time with exquisite subcellular resolution. However, FRET-based live-cell imaging requires application-specific biosensors and specialized imaging systems, limiting its use in studies. For complete details on the use and execution of this protocol, please refer to Socodato et al. (2020), Portugal et al. (2017), and Socodato et al. (2018).
本方案重点介绍了利用荧光共振能量转移（FRET）生物传感器实时研究小胶质细胞激活过程中的信号事件。由于小胶质细胞激活有助于解析神经退行性疾病中发生的神经病变过程的信号机制，因此其研究受到了广泛关注。与广泛应用于小胶质细胞领域的更传统方法不同，FRET 可实现对小胶质细胞信号事件的实时研究，并具有精细的亚细胞分辨率。然而，基于 FRET 的活细胞成像需要特定于应用的生物传感器和专门的成像系统，这限制了其在研究中的应用。如需详细了解本方案的使用和执行方法，请参考 Socodato 等人（2020 年）、Portugal 等人（2017 年）和 Socodato 等人（2018 年）的研究。
