中性粒细胞碱性磷酸酶染色实验条件探讨

Study on Experimental Conditions of Neutrophil Alkaline Phosphatase Staining

目的探讨中性粒细胞碱性磷酸酶染色的实验条件。方法用40%、60%、80%的丙酮-枸缘酸固定液对中性粒细胞碱性磷酸酶染色固定条件进行测定。同时用2-氨基-2-甲基-1.3-丙二醇(pH9.4-9.6)缓冲液、巴比妥(pH9.2)缓冲液、Tris(pH9.2)缓冲液配制的基质液分别在10、15、20分钟的染色条件进行测定。结果是选择60%的丙酮-枸缘酸固定30秒染色结果最好。三种不同的缓冲液和三个不同的孵育时间的染色结果经多因素方差分析,P<0.05,结果有统计学意义。结论最佳染色条件为60%的丙酮-枸缘酸固定30秒,用Tris(pH9.2)缓冲液配制的基质液,孵育时间为15分钟。本方法帮助鉴别慢性粒细胞性白血病和类白血病有较大的实用价值。

Objective To discuss the experimental conditions of neutrophil alkaline phosphatase staining. Methods The immobilizing conditions of neutrophil alkaline phosphatase staining were detected by the stationary liquid of 40%, 60% and 80% acetone - citric acid, and at the same time, the staining conditions were detected at 10, 15 and 20 minutes in the matrix liquid prepared respectively with 2 -a- mino - 2 - methyl - 1.3 - prepylene glycol （ PH9. 4 - 9.6 } buffer, barbital （ PH9. 2 ） buffer and riffs { PH9. 2 } buffer. Results Fixing in 60% acetone - citric acid for 30 seconds gave out the best staining results ; the staining results of the three different buffers and the three different incubating times received the multiple factor variance analysis, the differences among them was of statistical significance { P 〈 0. 05 }. Conclusions The best staining conditions include fixing in 60% acetone - citric acid for 30 seconds, the matrix liquid prepared with riffs （ PHg. 2 ） buffer and incubation for 15 minutes ; this method is of practical value in helping identify chronic myeloid leukemia and leukemoid reaction.