FIBROBLAST

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。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 研究中心，而研究中心不一定是研究者的研究机构。 我们已经基于边缘的形态学和运动学以及存在的蛋白质定义了在细胞边缘发生的许多不同类型的运动性（Dubin-Thaler，提交;（Dobereiner et al.，2005））。 由于前缘通常非常薄（200 nm），因此应该可以在冷冻电镜中观察前缘处复合物的分子水平形态。 许多复合物已在体外可视化，因此，它可能是可能的，以确定在体内的复合物，这将使一个更好地了解不同类型的运动在分子水平上。 特别是，我们建议准备胞质体（细胞质的去核部分）和纤连蛋白表面上的传播特性。 然后，当我们在光学显微镜下记录细胞质时，细胞质就会扩散。 这将使我们能够确定正在发生的运动类型。 在适当的时候，样品将用甲醛固定，随后将在非常薄的玻璃状冰层中冷冻。 或者，允许样品扩散一段规定的时间，然后冷冻而不固定。

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 have defined a number of different types of motility that occur at the edges of cells based upon the morphology and kinematics of the edge as well as the proteins present (Dubin-Thaler, submitted; (Dobereiner et al., 2005)). Because the leading edge is typically very thin (<200 nm), it should be possible to visualize the molecular-level morphology of the complexes at the leading edge in the cryo-em. Many of the complexes have been visualized in vitro; therefore, it may be possible to identify the in vivo complexes, which will enable a much better understanding of the different types of motility at the molecular level. In particular, we propose to prepare cytoplasts (enucleated portions of cytoplasm) and characterize their spreading on fibronectin surfaces. Cytoplasts would then be spread as we recorded them in the light microscope. This would enable us to define the type of motility that was occurring. At the appropriate time, the sample would be fixed with formaldehyde and would subsequently be frozen in a very thin layer of vitreous ice. Alternatively, the samples would be allowed to spread for a defined period and then frozen without fixation.