Magnetic flow sorting using a model system of human lymphocytes and a colloidal magnetic label.
Magnetic flow sorting using a model system of human lymphocytes and a colloidal magnetic label.
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使用人类淋巴细胞模型系统和胶体磁性标记进行磁流分选。
DOI：
10.1097/00002480-199609000-00071
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Chalmers,JJ
中科院分区：
文献类型：
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作者：
Zborowski,M;Moore,LR;Reddy,S;Chen,GH;Sun,L;Chalmers,JJ
具有相同物理性质但表面蛋白表达不同的细胞可以使用与荧光或磁性标记缀合的免疫特异性染色剂方便地进行分选。使用商业批量分选机的免疫磁性细胞分选提供了高分选能力、分选组分的高活力和高消耗率的优点;其缺点是低富集率和批量处理。作者开发并测试了用于小体积实验性细胞富集的连续、流通式磁性细胞分选仪。新鲜分离的人外周血淋巴细胞标记使用免疫荧光磁性夹心组成的小鼠抗人CD 8单克隆抗体-荧光素缀合物和大鼠抗小鼠多克隆抗体-胶体铁-葡聚糖缀合物。使用1.334 T的饱和磁场和五通道分选仪每小时分选总共2-3百万个淋巴细胞。荧光细胞分布在通道之间，与它们的荧光强度和磁化率有关。纯度（68-85%）和富集率（16- 34倍）与商业批量磁选机相当;分选能力和富集馏分的回收率（高达32%）受到分选机小规模的限制。未来的发展方向是提高富集组分的分辨率、回收率和分选能力，并在其他细胞系统上测试分选仪。
Cells of identical physical properties that differ in the expression of surface proteins can be sorted conveniently using immunospecific stains conjugated to fluorescent, or magnetic, labels. Immunomagnetic cell sorting using commercial batch sorters offers advantages of high sorting capacity, high viability of sorted fractions, and high depletion rates; its disadvantages are low enrichment rate and batch processing. The authors developed and tested a continuous, flow-through magnetic cell sorter for small volume, experimental cell enrichment. Freshly isolated human peripheral lymphocytes were labeled using an immunofluoromagnetic sandwich consisting of mouse anti-human CD8 monoclonal antibody-fluorescein conjugate and rat anti mouse polyclonal antibody-colloidal iron-dextran conjugate. A total of 2–3 mln lymphocytes were sorted per hour using a saturation magnetic field of 1.334 T and a five channel sorter. The fluorescent cells were distributed among the channels in relation to their fluorescence intensity and magnetic susceptibility. The purity (68–85%) and enrichment rates (16–34X) were comparable to those of commercial batch magnetic separators; sorting capacity and recovery of the enriched fractions (up to 32%) were limited by the small scale of the sorter. Future direction is focused on increasing the resolution, recovery, and sorting capacity of the enriched fractions, and testing the sorter on other cell systems.