制备豚鼠补体时避免溶血发生的方法学探讨

Methods of Avoiding Hemolysis in Preparing Serum Complements of Guinea Pigs

[目的]研究制备豚鼠补体时避免溶血发生的方法。[方法]方法1为先混合8只豚鼠的血清,再将绵羊红细胞(SRBC)加入到混合的豚鼠血清中进行吸附;方法2为先把SRBC分别加入到豚鼠血清中进行吸附,再将吸附后的8只豚鼠血清混合,然后肉眼观察2种处理方法所得样品的溶血现象,并在540nm波长下测定其吸光度(D)值。[结果]按方法1所制备的豚鼠补体,肉眼可见明显溶血现象;而采用方法2则溶血现象不明显,且方法2组样品吸光度值低于方法1组(P<0.05)。[结论]采用先吸附,后混合的方法,可较好地避免豚鼠补体制备时发生溶血。

[Objective] To explore the methods of avoiding hemolysis in preparing serum complements of guinea pigs. [Methods] In method 1, the serum of 8 guinea pigs was fixed first and then sheep red blood cells (SRBC) were added to the mixed serum for absorption. In method 2, SRBC was added to the serum of 8 guinea pigs respectively for absorption first and then the serum containing SRBC was mixed. After the experiment, macroscopic examination was carried out to observe the hemolysis and the optical density of the two kinds of mixture was detected at the wavelength of 540 nm. [ Results] In the serum complements of guinea pigs prepared by method 1, macroscopic hemolysis occurred; in the serum complements prepared by method 2, macroscopic hemolysis was not obvious and the optical density of the mixture was lower than that by method 1 ( P < 0.05) . [Conclusion] In preparing the serum complements of guinea pigs, the method of 'mixing following absorption' is better for the avoidance of hemolysis in preparing serum complements of guinea pigs.