MOBILITY STUDY OF ALLYLAMINE BY FRAP

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。我们感兴趣的是研究流动性的表面吸附的聚烯丙基胺使用荧光恢复后光漂白（FRAP），单分子为基础的技术。这项技术使我们能够直接测量细胞膜中蛋白质和脂质的横向运输速率。对我们来说，最感兴趣的三个问题是：（1）聚（烯丙基胺）是否在任何条件下从逐层表面[与聚（苯乙烯磺酸盐）底层]脱落;例如，酸、碱、高盐浓度，（2）在任何条件下外部聚（烯丙胺）层是否存在任何横向迁移性;即，FRAP在这个外层，以及（3）如何我们的新的共价交联聚（烯丙基胺）表面相比，传统的离子交联，逐层表面的稳定性和横向流动性。为此，聚烯丙基胺用荧光染料标记，该荧光染料可以由氩离子激光器的514 nm线激发，氩离子激光器又耦合到共聚焦显微镜。

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. We are interested to study the mobility of surface adsorbed polyallylamine using Fluorescence Recovery After Photobleaching (FRAP), a single molecule based technique. This technique gives us a direct measure of the rates of lateral transport of proteins and lipids in cell membranes. For us, the three issues of greatest interest are: (1) Does poly(allylamine) come off of a layer-by-layer surface [with an underlayer of poly(styrenesulfonate)] under any conditions; e.g., acid, base high salt concentration, (2) is there any lateral mobility of the outer poly(allyamine) layer under any conditions; i.e., FRAP within this outer layer, and (3) how does our novel covalently-cross-linked poly(allylamine) surface compare with conventional ionically-cross-linked, layer-by-layer surfaces in terms of stability and lateral mobility. For this the polyallylamine is labeled with a fluorescent dye that can be excited by the 514 nm line of an Argon-ion laser which in turn is coupled to a confocal microscope.