染色体畸变分析实验条件探讨

目的探讨如何确立外周血淋巴细胞培养染色体畸变分析实验条件,以获得良好的制片效果。方法在已消毒好备用的超净台内,用碘伏消毒采血管的头部,轻轻混匀后接种,加入秋水仙素工作液,经混匀、培养、收集细胞并离心后,弃上清液,加入低渗液混匀,给予预固定;重复固定后依细胞数量加入适量固定液,制片、染色、气干。于低倍镜下细致观察,计数200个分裂相。结果细胞生长良好,分裂指数高,能够满足分析要求,染色体分散均匀,着色良好,长短适中,两条姐妹染色体大致平行分离,着丝粒清晰。结论寻找适合自己实验室的实验条件能提高染色体畸变的检出率和阅片速度,及时准确地反映辐射损伤情况,对放射工作人员健康进行监护,为意外放射性损伤事故进行生物剂量估算,为放射患者诊断与治疗提供依据。

Investigation on experimental conditions of chromosome aberration

Objective To investigate the experiment condition of peripheral blood lymphocyte culture for chromosome aberration analysis to obtain a good production effect. Methods In the sterile spare super clean bench, after disinfecting the needle, inoculating in the mixed culture medium, taking colchicine solution into the mixture, the culture began. Matured cells, which were collected and centrifuged, with discarding the supernatant and adding hypotonic liquid, were pre-fixed. With the repeated fixation, cells were fixed with stationary liquid, then taken into produced chromosome slides, after dyeing and air drying. Observed at low magnification, 200 mitotic cells were counted. Results Cell growth was good with high mitotic index, which met the analytic need. The dispersion, color and length of chromosome were good, and sister chromatid separation roughly paralleled, with clear centromere. Conclusion Suitable experimental conditions can make the improvement of the detection rate of chromosome aberration and the speed of reading slides, which can reflect the radiation damage timely and accurately, take care the health of radiation workers, estimate the biological dose of radiation accident to provide the basis of the diagnosis and treatment for the patient.