Ellett Felix, Lieschke Graham J
Australian Regenerative Medicine Institute, Monash University, Clayton, Victoria, Australia.
Methods Enzymol. 2012;506:425-35. doi: 10.1016/B978-0-12-391856-7.00046-9.
Fluorescent transgenes with leukocyte-restricted expression have been essential to recent studies using zebrafish models of inflammation and the innate immune response. Many of the experiments performed using these models involve quantifying changes in the number of fluorescent leukocytes. Here, we describe a tool for deriving a quantitative variable proportional to fluorescent leukocyte numbers from single-plane fluorescent digital images of whole live embryos. The parameter, called "Leukocyte units," provides reliable values linearly proportional to actual leukocyte numbers in the range 50-400 leukocytes/embryo, and its performance at higher leukocyte densities remains linear. Its usefulness is demonstrated by scoring changes in leukocyte numbers following perturbation of a characterized leukocyte specification pathway.
具有白细胞限制性表达的荧光转基因对于最近使用斑马鱼炎症模型和先天免疫反应模型的研究至关重要。使用这些模型进行的许多实验都涉及量化荧光白细胞数量的变化。在这里，我们描述了一种从整个活胚胎的单平面荧光数字图像中得出与荧光白细胞数量成比例的定量变量的工具。这个参数称为“白细胞单位”，在50 - 400个白细胞/胚胎的范围内提供与实际白细胞数量成线性比例的可靠值，并且在更高白细胞密度下其性能仍保持线性。通过对一个特定白细胞分化途径受到干扰后白细胞数量的变化进行评分，证明了它的实用性。
