流式细胞术检测圆鳍鱼白细胞的胞吞功能和呼吸爆发活性

niwanmao

这是一篇鱼类的研究文献，发表在2012年十月份的PLOS ONE上，主要是学习一下他们的胞吞功能检测（Fluoresbrite YG carboxylate microspheres的吞噬）和呼吸爆发活性的检测（dihydrorhodamine 123 (DHR）被氧化为罗丹明123），方法学见文献的第5页（http://www.flowcyto.cn/bbs/reade ... nHw8I9Ut0ohPKyC8NJI）。结果图片如下：

胞吞作用

呼吸爆发

Phagocytosis and Respiratory Burst Activity in Lumpsucker (Cyclopterus lumpus L.) Leucocytes Analysed by Flow Cytometry
Gyri T. Haugland, Ragnhild Aakre Jakobsen, Nils Vestvik, Kristian Ulven, Lene Stokka, Heidrun I. Wergeland

Abstract
In the present study, we have isolated leucocytes from peripheral blood, head kidney and spleen from lumpsucker (Cyclopterus lumpus L.), and performed functional studies like phagocytosis and respiratory burst, as well as morphological and cytochemical analyses. Different leucocytes were identified, such as lymphocytes, monocytes/macrophages and polymorphonuclear cells with bean shaped or bilobed nuclei. In addition, cells with similar morphology as described for dendritic cells in trout were abundant among the isolated leucocytes. Flow cytometry was successfully used for measuring phagocytosis and respiratory burst activity. The phagocytic capacity and ability were very high, and cells with different morphology in all three leucocyte preparations phagocytised beads rapidly. Due to lack of available cell markers, the identity of the phagocytic cells could not be determined. The potent non-specific phagocytosis was in accordance with a high number of cells positive for myeloperoxidase, an enzyme involved in oxygen-dependent killing mechanism present in phagocytic cells. Further, high respiratory burst activity was present in the leucocytes samples, verifying a potent oxygendependent degradation. At present, the specific antibody immune response could not be measured, as immunoglobulin or B-cells have not yet been isolated. Therefore, analyses of the specific immune response in this fish species await further clarification. The present study presents the first analyses of lumpsucker immunity and also the first within the order Scopaeniformes.


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