Assas Bakri M, Abdulaal Wesam H, Wakid Majed H, Zakai Haytham A, Miyan J, Pennock J L
Faculty of Biology Medicine and Health, University of Manchester, UK; Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, King AbdulAziz University, Jeddah, Saudi Arabia.
Department of Biochemistry, Cancer Metabolism and Epigenetic Unit, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia.
Anal Biochem. 2017 Jun 15;527:13-19. doi: 10.1016/j.ab.2017.03.025. Epub 2017 Mar 31.
Flow cytometric analysis of calcium mobilisation has been in use for many years in the study of specific receptor engagement or isolated cell:cell communication. However, calcium mobilisation/signaling is key to many cell functions including apoptosis, mobility and immune responses. Here we combine multiplex surface staining of whole spleen with Indo-1 AM to visualise calcium mobilisation and examine calcium signaling in a mixed immune cell culture over time. We demonstrate responses to a TRPV1 agonist in distinct cell subtypes without the need for cell separation. Multi parameter staining alongside Indo-1 AM to demonstrate calcium mobilization allows the study of real time calcium signaling in a complex environment.
钙动员的流式细胞术分析已在特定受体结合或分离细胞间通讯的研究中使用多年。然而，钙动员/信号传导是许多细胞功能的关键，包括细胞凋亡、迁移和免疫反应。在此，我们将全脾的多重表面染色与Indo-1 AM相结合，以可视化钙动员，并随时间检查混合免疫细胞培养物中的钙信号传导。我们证明了在无需细胞分离的情况下，不同细胞亚型对TRPV1激动剂的反应。与Indo-1 AM一起进行多参数染色以证明钙动员，能够在复杂环境中研究实时钙信号传导。
