Calibrated Particles for Cell Adhesion Assays

9712656 Quinn许多重要的微生物和细胞过程需要了解这些实体附着在表面、其他细胞和受体蛋白质上的作用力。其中一些过程包括细胞分离、肿瘤转移和壁贴壁、血管假体的细胞种植等。大多数黏附分析本质上是非常定性的，通常是由于容器“血管”的不规则性质、其尺寸小，以及在没有对设备开发的广泛投资的情况下很难获得定量数据。粘附力研究通常是通过测量将细胞从底物上洗掉的力来完成的。这项SGER建议试图通过开发一组已知表面受体强度的细胞来校准新的测量血管和细胞受体系统，从而使这一过程更加简单和定量。标准细胞将是乳胶珠，它已经被涂上了用于实验的适当的受体，并标有识别标签。涂覆的乳胶颗粒的粘附性将在径向流动分离分析(RFDA)装置中确定，该装置是PI和同事在先前资助的基础上开发的。已知粘附力(抗剪切洗涤力)的细胞然后可以用来更准确地确定简单洗涤测试中所涉及的力，例如，用移液管水流从涂覆的井中洗涤细胞。***

9712656 Quinn Many important microbial and cellular processes require a knowledge of the forces with which these entities adhere to surfaces, other cells, and receptor proteins. some of these processes include cell separations, tumor metastasis and wall adhesion, cell seeding for vascular prostheses, etc. Most adhesion assays are very qualitative in nature, frequently due to the irregular nature of the containment "vessels", their small size, and the difficulty in obtaining quantitative data without extensive investment in device development. Adhesion studies are usually done by measuring the force to wash the cell off of the substrate. This SGER proposal seeks to make the process simpler and more quantitative by developing a set of cells of known surface-receptor strength for use in calibrating new measurement vessels and cell receptor systems. The standard cells will be latex beads which have been coated with the appropriate receptor for the experiment and labeled with an identifying tag. The adhesion of the coated latex particles will be determined in the Radial Flow Detachment Assay (RFDA) device that the PI and coworkers have developed over the previous funded grant. The cells of known adhesion (resistance to shear washing force) could then be used to determine more accurately the forces involved in simple washing tests, e.g., washing of cells from coated wells by pipette streams of water. ***