HUMAN PMNL MEMBRANE MICRODOMAINS AND THEIR MODIFICATIONS BY DEHP

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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.