Siiman O, Gordon K, Burshteyn A, Maples J A, Whitesell J K
Advanced Technology, Beckman Coulter, Miami, Florida 33196-2500, USA.
Cytometry. 2000 Dec 1;41(4):298-307. doi: 10.1002/1097-0320(20001201)41:4<298::aid-cyto8>3.0.co;2-0.
The type of antibody-conjugated polystyrene (PS) latex beads for use as light scatter shift agents for targeted lymphocyte populations in whole blood has been expanded to include gold and silver nanoparticle-aminodextran-PS latex bead conjugates with antibodies. The linkers between antibody and colloidal metal were an aminotrithiol ligand or aminodextran polymer molecules.
A modified flow instrument, including forward light scatter (FS), side light scatter (SS), light scatter at other intermediate angle ranges, LMALS (10-20 degrees ) and UMALS (20-65 degrees ) was used for simultaneous bead probe measurements. A conventional flow cytometer was used in simultaneous bead-fluorescent marker experiments.
Two mutually exclusive cell populations, CD4+ and CD8+ lymphocytes, have been simultaneously enumerated in blood by using a mixture of CD4-PS, CD8-Au-PS or CD4-Au-PS, CD8-PS beads, and one laser line, 633 nm, excitation. Similar measurements were made with mixtures of CD4-PS, CD8-Ag-PS or CD4-Ag-PS, CD8-PS beads. Also, simultaneous use of bead and fluorescent markers mixed with whole blood was demonstrated with CD4-PS beads and with the CD4-RD1/CD8-FITC dual marker.
Enumeration of CD4 and CD8 lymphocytes in whole blood by light scatter parameters only compared well with standard analyses with fluorescent markers. In simultaneous bead-fluorescent marker labeling of lymphocytes, the labeled bead had to be mixed first with cells in whole blood.
用作全血中靶向淋巴细胞群体光散射转移剂的抗体偶联聚苯乙烯（PS）乳胶珠类型已扩展到包括与抗体结合的金和银纳米颗粒 - 氨基葡聚糖 - PS乳胶珠缀合物。抗体与胶体金属之间的连接体是氨基三硫醇配体或氨基葡聚糖聚合物分子。
使用一种改良的流式仪器，包括前向光散射（FS）、侧向光散射（SS）、其他中间角度范围的光散射、大角度多角度光散射（LMALS，10 - 20度）和小角度多角度光散射（UMALS，20 - 65度）进行同时的珠探针测量。在同时进行珠 - 荧光标记实验时使用传统流式细胞仪。
通过使用CD4 - PS、CD8 - Au - PS或CD4 - Au - PS、CD8 - PS珠的混合物以及一条激光线（633 nm激发），已在血液中同时计数了两个相互排斥的细胞群体，即CD4 +和CD8 +淋巴细胞。使用CD4 - PS、CD8 - Ag - PS或CD4 - Ag - PS、CD8 - PS珠的混合物进行了类似测量。此外，还证明了将珠与荧光标记物与全血混合同时使用，如CD4 - PS珠以及CD4 - RD1/CD8 - FITC双标记物。
仅通过光散射参数对全血中CD4和CD8淋巴细胞进行计数与使用荧光标记物的标准分析结果相当。在淋巴细胞的珠 - 荧光标记物同时标记中，标记的珠必须首先与全血中的细胞混合。
