Vaporciyan A A, Jones M L, Ward P A
Department of Pathology, University of Michigan Medical School, Ann Arbor 48109-0602.
J Immunol Methods. 1993 Feb 26;159(1-2):93-100. doi: 10.1016/0022-1759(93)90145-w.
Neutrophil adherence to endothelial monolayers in multi-well plates can be rapidly quantitated, using a fluorescent plate reader. The number of adherent neutrophils which have been labeled with biscarboxyethyl carboxyfluorescein (BCECF) was calculated by comparing fluorescence of endothelial monolayers to a reference standard curve determined by fluorescence intensity of suspensions of labeled neutrophils. The limit of sensitivity approached 100 cells, which is similar to that achieved by the most sensitive of alternate techniques. The assay, after isolation of the neutrophils, can be completed in 1.5 h. The ability to utilize microwell plates permits the use of large numbers of samples. This method has the advantage of being relatively simple and rapid while maintaining a sensitivity comparable to alternate approaches which employ radioactive labeling or counting of adherent or non-adherent cells.
使用荧光酶标仪可快速定量多孔板中中性粒细胞与内皮细胞单层的黏附情况。已用双羧乙基羧基荧光素（BCECF）标记的黏附中性粒细胞数量，是通过将内皮细胞单层的荧光与由标记中性粒细胞悬浮液的荧光强度确定的参考标准曲线进行比较来计算的。灵敏度极限接近100个细胞，这与最灵敏的其他技术所达到的灵敏度相似。中性粒细胞分离后，该检测可在1.5小时内完成。使用微孔板的能力允许检测大量样本。该方法的优点是相对简单、快速，同时保持与采用放射性标记或对黏附或未黏附细胞进行计数的其他方法相当的灵敏度。
