用三色间接流式细胞术检测猪外周血白细胞亚群

To Detect Swine Peripheral Blood Leukocytes by Indirect Immunostaining Method for Triple-color Flow Cytometry

用三色间接流式细胞术检测猪外周血粒细胞、B细胞和单核细胞亚群。来源于2头猪的12个猪外周血液样品裂解红细胞后,用3种SWC(swine workshop cluster)单抗和不同标记的二抗染色,对全体细胞设门,测定了SWC1、SWC3、SWC8在外周血白细胞上的表达,分析了嗜中性粒细胞(SWC1+SWC3+SWC8+)、嗜酸性粒细胞(SWC1-SWC3+SWC8+)、B细胞(SWC1-SWC3-SWC8+)、单核细胞(SWC1+SWC3+SWC8-)在外周血白细胞中的比例。结果显示,细胞分群情况良好,两组测定结果的均值及方差无显著差异,其中嗜中性粒细胞占(32.6±0.8)%,嗜酸性粒细胞占(4.7±0.3)%,B淋巴细胞占(3.7±0.2)%,单核细胞占(2.2±0.2)%。

The experiments were carried out to use an indirect immunostaining method for triple - color flow cytometry （FCM） to analyze swine peripheral blood leukocytes （PBL）. After erythrocytes lysated, 12 samples from 2 piglets were stained with three different primary and labeled secondary antibodies. The leukocyte subpopulations were identified using a SWC1/SWC3/SWC8 triple immunofluoreseence analysis with an electronic gate set on all PBL. The subpopulations of PBL were nicely detected ： neutrophil （ SWC1^＋ SWC3^＋ SWC8^＋ ） occupies （32.6 ± 0. 8） % of total PBL, eosinophil （ SWC1^- SWC3^＋ SWC8^＋） occupies （4.7± 0. 3 ） %, B cell （ SWC1^ - SWC3 ^- SWC8 ^＋ ） occupies （3.7 ± 0.2） % and monocyte （ SWC1^ ＋ SWC3 ^＋ SWC8^ - ） occupies （2.2 ± 0.2） %. Further statistics research indicated that average and variance of two groups of data did not showed significant difference.