Sun J T, Armstrong M J, Galloway S M
Merck Research Laboratories, W 45-3, West Point, PA 19486, USA.
Mutat Res. 1999 Feb 2;439(1):121-6. doi: 10.1016/s1383-5718(98)00175-2.
Micronuclei are routinely scored in anucleate erythrocytes in bone marrow smears stained with acridine orange. Intense fluorescence from the many nucleated cells in the preparations can interfere with micronucleus detection and cause fatigue in the reader. A method for removing nucleated cells by filtering bone marrow through cellulose packed in syringes was developed by Romagna some ten years ago, but has not been used routinely because of the excessive time needed to prepare columns. We have modified the method very simply by filling chromatography columns by pipet with a cellulose suspension. We show here that column filtration of bone marrow does not affect the numbers of micronucleated polychromatic erythrocytes (MN-PCEs) scored from mice treated with the chromosome breaking agents mitomycin C and cyclophosphamide, or the aneuploidy-inducing spindle poisons, colchicine and vinblastine. The extra preparation time is only about half an hour for a full scale micronucleus assay, and results in better slides and faster scoring.
在经吖啶橙染色的骨髓涂片的无核红细胞中常规对微核进行计数。制备物中众多有核细胞发出的强烈荧光会干扰微核检测，并使阅片者感到疲劳。大约十年前，罗马涅开发了一种通过用装在注射器中的纤维素过滤骨髓来去除有核细胞的方法，但由于制备柱所需时间过长，尚未常规使用。我们通过用移液管将纤维素悬浮液填充到色谱柱中，非常简单地改进了该方法。我们在此表明，骨髓的柱过滤不会影响用染色体断裂剂丝裂霉素C和环磷酰胺或诱导非整倍体的纺锤体毒物秋水仙碱和长春碱处理的小鼠中计数的微核多染性红细胞（MN - PCE）的数量。对于全面的微核试验，额外的制备时间仅约半小时，并且能得到更好的玻片并加快计数速度。
