Satake Hiroto, Sakata Toshiya
Department of Materials Engineering, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
ACS Omega. 2019 Aug 22;4(10):14255-14260. doi: 10.1021/acsomega.9b01872. eCollection 2019 Sep 3.
In this paper, we clarify the interfacial pH behavior induced by allergic responses at a mast cell/gate insulator nanogap detected by laser scanning confocal fluorescence microscopy. In a previous work, the change in interfacial pH detected on the basis of allergic responses was monitored at a mast cell/gate insulator nanogap interface using a cell-cultured gate ion-sensitive field-effect transistor (ISFET), but the interfacial pH behavior at a mast cell/gate insulator nanogap has not been clarified using other methods. Here, the phospholipid fluorescein is employed as the extracellular pH indicator, which is fixed to the external side of the plasma membrane of mast cells cultured on a substrate. As a result, the interfacial pH at the mast cell/substrate nanogap increases after mast cells with IgE on their membrane are activated by the interaction between IgE and an allergen. This is due to the basicity of histamine molecules released from mast cells. Moreover, the change in the interfacial pH at the mast cell/substrate nanogap is larger than that at the mast cell/bulk solution interface. That is, molecules of substances secreted as a result of allergic responses are assumed to accumulate around the cell/substrate nanogap. The data obtained in this study support the idea that potentiometric ion sensors such as ISFETs can detect a cellular-function-induced change in pH at a cell/electrode nanogap in real time.
在本文中,我们阐明了通过激光扫描共聚焦荧光显微镜检测到的肥大细胞/栅极绝缘体纳米间隙处过敏反应诱导的界面pH行为。在先前的工作中,使用细胞培养的栅极离子敏感场效应晶体管(ISFET)在肥大细胞/栅极绝缘体纳米间隙界面监测基于过敏反应检测到的界面pH变化,但尚未使用其他方法阐明肥大细胞/栅极绝缘体纳米间隙处的界面pH行为。在此,磷脂荧光素被用作细胞外pH指示剂,其固定在培养在底物上的肥大细胞质膜的外侧。结果,膜上带有IgE的肥大细胞在IgE与过敏原相互作用后被激活,肥大细胞/底物纳米间隙处的界面pH升高。这是由于肥大细胞释放的组胺分子呈碱性。此外,肥大细胞/底物纳米间隙处的界面pH变化大于肥大细胞/本体溶液界面处的变化。也就是说,假定过敏反应分泌的物质分子在细胞/底物纳米间隙周围积累。本研究获得的数据支持这样的观点,即诸如ISFET之类的电位离子传感器可以实时检测细胞功能诱导的细胞/电极纳米间隙处的pH变化。