Phenotypic analysis using very small volumes of blood.
Phenotypic analysis using very small volumes of blood.
复制标题
使用极少量的血液进行表型分析。
DOI：
10.1002/0471142956.cy0630s54
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
Germolec,DoriR
中科院分区：
文献类型：
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作者：
Weaver,JamesL;McKinnon,Katherine;Germolec,DoriR
分析外周血白细胞的细胞表面表型是流式细胞术最常见的应用之一。在老鼠的研究中，动物的小尺寸限制了可用的血量。使用红细胞裂解或梯度分离然后反复洗涤的标准染色方法涉及不可避免的细胞损失，这通常限制了对血液的分析到最终方法。因此，时间进程研究需要在每个时间点牺牲一组小鼠。因此，需要一种可以与更少的血量一起使用的方法。这将允许随着时间的推移对同一动物进行连续采样，减少实验变异性，减少动物使用。这里描述的方法是不裂解、不洗涤的方法，它使用对荧光参数的触发。该方法允许使用每管20微米L的全血容量进行常规的外周血白细胞表型分析。这些数据可以与需要更多血量的传统方法得出的值相比较。由于红细胞的干扰，光散射参数在这种方法中是不可用的。这种方法已经被用于持续长达四周的小鼠外周血细胞数量的时程研究。普罗托克。赛姆。54：6.30.1-6.30.8。2010John Wiley&Sons，Inc.
Analysis of cell‐surface phenotype of peripheral blood leukocytes is one of the most common applications of flow cytometry. In mouse research, the small size of the animal limits the amount of blood available. Standard staining methods using lysis of erythrocytes or gradient separation followed by repeated washing involve unavoidable losses of cells that generally limit analysis of blood to terminal methods. Time‐course studies, therefore, require sacrifice of groups of mice at each time point. Thus, a method is needed that can be used with much smaller volumes of blood. This will allow serial sampling of the same animal over time, decreasing experimental variability and reducing animal use. The method described here is a no‐lyse, no‐wash method that uses triggering on a fluorescence parameter. The method allows routine analysis of the phenotype of peripheral blood leukocytes using whole‐blood volumes of 20 µl per tube. The data are comparable with values from traditional methods requiring much higher volumes of blood. Due to interference by erythrocytes, light‐scatter parameters are not usable with this method. This method has been used for time‐course studies of peripheral blood populations in mice lasting as long as four weeks.Curr. Protoc. Cytom. 54:6.30.1‐6.30.8. © 2010 by John Wiley & Sons, Inc.