Centro de Biología Molecular Severo Ochoa, Consejo Superior de investigaciones Científicas, Universidad Autónoma de Madrid, Madrid, Spain.
Methods Mol Biol. 2023;2675:109-115. doi: 10.1007/978-1-0716-3247-5_9.
The study of immunometabolism is an important and emerging field in immunology. B-cell activation upon antigen recognition induces profound metabolic changes in the cell, leading to an increase in ATP production to sustain cell proliferation and differentiation. Current methods available to determine the amount of ATP are time-consuming, require extensive sample processing, and need a large amount of starting material. We set up an easy follow-up protocol to determine the relative amount of ATP in living cells, combining cell surface staining with quinacrine. This acridine dye emits a green fluorescent signal in the presence of intracellular ATP. This protocol allows us to determine ATP in small populations of cells using flow cytometry, such as the germinal center.
免疫代谢研究是免疫学中一个重要且新兴的领域。B 细胞在抗原识别后会引起细胞内代谢的深刻变化，导致 ATP 产生增加，以维持细胞增殖和分化。目前用于测定 ATP 含量的方法耗时耗力，需要对样本进行大量处理，并且需要大量的起始材料。我们建立了一种简单的后续方案来确定活细胞中相对的 ATP 含量，将细胞表面染色与吖啶橙结合起来。这种吖啶染料在存在细胞内 ATP 的情况下会发出绿色荧光信号。该方案允许我们使用流式细胞术来确定如生发中心等小细胞群中的 ATP。
