CELL SELECTION BY MAGNETIC FLOW SORTING

描述：这项提议将开发一种磁性细胞分选方法 用于连续流动中的阳性和阴性细胞选择。结果是 前一个供资期间支持这一做法。高产量、高产量 高纯磁选方法将产生重大的积极影响 在细胞生物学和其他应用方面的研究。申请者有 选择了两种有希望的设计：偶极和四极磁选机， 和两种很有希望的可溶性磁性标记：胶体铁葡聚糖和 分子磁铁蛋白。他们开发了用于运动分析的工具 使用粒子跟踪测速仪(PTV)测量磁场和流场。 具体目标是(1)优化磁选机的性能。两个人 分拣机将进行优化，以在最大体积上获得最大磁力 样本中的。将评估流体动力聚焦在 偶极分离器。将使用场流分馏来优化 四极分离器的性能。磁力的影响 与细胞表面抗原密度有关的敏感性分布 分选馏分的回收率和纯度将由人类进行研究 外周血淋巴细胞。新型参考磁化率珠基 将开发一种用于标准化分拣机的动态分选器。最好的 执行分选机将扩大规模，用于临床样本。这个 第二个目标是确定细胞磁场的分布 PTV的易感性。之前的研究结果表明，这种偏差 从高场下细胞磁矩与场之间的线性关系 梯度导致了细胞磁化率测量的较大误差。更高 B对晶胞磁化的有序贡献将被分析以增加 测量的精度。细胞易感频率 将测量细胞亚型内部和之间的铁分布 葡聚糖和磁铁蛋白免疫磁标记物，并与 流式细胞术检测抗原密度分布。目标3将测试 磁流分选机在大单元体积上的性能。原型机将 检测外周血单核细胞中干细胞的浓缩情况 细胞。

DESCRIPTION: This proposal will develop a method of magnetic cell sorting for positive and negative cell selection in a continuous flow. Results in the previous funding period support this approach. A high yield, high purity magnetic separation method would have a significant positive impact on research in cell biology and other applications. The applicants have selected two promising designs: dipole and quadrupole magnetic separators, and two promising soluble magnetic labels: colloidal iron dextran and molecular magnetoferritin. They have developed tools for motion analysis in the magnetic and flow fields, using particle tracking velocimetry (PTV). The specific aims are (1) To optimize magnetic sorter performance. The two sorters will be optimized for maximum magnetic force over the largest volume of the sample. Hydrodynamic focusing will be evaluated for use in the dipole separator. Field-flow fractionation will be used to optimize performance of the quadrupole separator. The effects of magnetic susceptibility distribution, related to cell surface antigen density, on the recovery and purity of sorted fractions will be studied with human peripheral lymphocytes. Novel reference magnetic susceptibility beads based on Dynabeads will be developed to standardize the sorter. The best performing sorter will be scaled up for use with clinical samples. The second aim is to determine the distribution of cell magnetic susceptibilities by PTV. Results of previous studies suggest the deviation from linearity between the cell magnetic moment and field at high field gradients leads to large error in cell susceptibility measurement. Higher order contributions of B to cell magnetization will be analyzed to increase the precision of the measurement. Cell susceptibility frequency distributions within and between cell subtypes will be measured for iron dextran and magnetoferritin immunomagnetic labels, and compared to the antigen density distribution by flow cytometry. Aim 3 will test the magnetic flow sorter performance on large cell volumes. The prototype will be tested for enrichment of stem cells from peripheral blood mononuclear cells.