HUMAN PMNL MEMBRANE MICRODOMAINS AND THEIR MODIFICATIONS BY DEHP

该子项目是利用 NIH/NCRR 资助的中心拨款提供的资源的众多研究子项目之一。子项目和研究者 (PI) 可能已从另一个 NIH 来源获得主要资金，因此可以在其他 CRISP 条目中出现。列出的机构是中心的机构，不一定是研究者的机构。之前使用 Laurdan 标记的 PMNL 悬浮液的稳态荧光进行的研究表明，塑料软化剂 DEHP 可以增加 PMNL 膜的流动性。特别是，该化合物降低了刚性较大的膜域的有序参数。本研究的目的是通过使用双光子显微镜观察单细胞的膜流动性域，然后研究塑料软化剂 DEHP 如何影响它们。 PMNL 将通过 Percoll 梯度离心从人血液中纯化，孵育或不孵育 DEHP，使其沉降到玻璃盖玻片上，用 Laurdan 标记，并在 37°C 下通过双光子显微镜分析 Laurdan GP。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Previous work using steady-state fluorescence of Laurdan-labelled PMNL suspensions showed that the plastic softener DEHP increases PMNL membrane fluidity. In particular, the compound decreases the order parameter of the more rigid membrane domains. The aim of this study is to visualize membrane fluidity domains on single cells by using two-photon microscopy and then investigate how the plastic softener DEHP may affect them. PMNL will be purified from human blood by Percoll gradient centrifugation, incubated or not with DEHP, allowed to settle onto a glass coverslip, labelled with Laurdan and analysed for Laurdan GP by two-photon microscopy at 37¿C.