CELL SELECTION BY MAGNETIC FLOW SORTING

描述：该提案将开发一种磁性细胞分选方法 用于连续流中的阳性和阴性细胞选择。 导致 上一个供资期支持这一做法。 高产量，高 纯度磁分离方法将有显着的积极影响 细胞生物学和其他应用的研究。 申请人已经 选择了两种有前途的设计：偶极和四极磁分离器， 和两种有前途的可溶性磁性标记：胶体铁葡聚糖和 分子磁铁蛋白 他们开发了运动分析工具， 磁场和流场，使用粒子跟踪测速（PTV）。 具体目标是：（1）优化磁分选机性能。 两 分拣机将优化最大磁力在最大的体积 的样本。 将对流体动力学聚焦进行评价， 偶极分离器 场流分离将用于优化 四极分离器的性能。 磁场的影响 敏感性分布，与细胞表面抗原密度有关， 回收率和纯度的分选馏分将研究与人类 外周血淋巴细胞 基于新型参考磁化率珠的 将开发Dynabeads上的产品，以标准化分拣机。 最好的 将按比例放大执行分选机以用于临床样本。 的 第二个目的是确定细胞磁性的分布 PTV的可接受性。 先前的研究结果表明， 从高场下细胞磁矩和场之间的线性 梯度导致细胞磁化率测量中的大误差。 更高 将分析B对单元磁化的阶次贡献以增加 测量的精度。 细胞易感性频率 将测量细胞亚型内和细胞亚型之间的铁分布 葡聚糖和磁铁蛋白免疫磁性标记，并与 通过流式细胞术测定抗原密度分布。 目标3将测试 大细胞体积上的磁流分选器性能。 原型将 检测从外周血单核细胞中富集干细胞 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.