Tamizh Selvan G, Chaudhury N K, Venkatachalam P
Sri Ramachandra University, Department of Human Genetics, Porur, Chennai 600 116, Tamil Nadu, India; Institute of Nuclear Medicine and Allied Sciences, Timarpur, New Delhi 110 054, India.
Institute of Nuclear Medicine and Allied Sciences, Timarpur, New Delhi 110 054, India.
Appl Radiat Isot. 2015 Mar;97:70-77. doi: 10.1016/j.apradiso.2014.12.018. Epub 2014 Dec 19.
Scoring micronuclei in the peripheral blood lymphocytes of individuals exposed to ionizing radiation is a rapid biodosimetry assay. Peripheral blood lymphocytes from five individuals were exposed in vitro to 0-5Gy of (60)Co γ-radiation at a dose rate of 0.76Gy/min. The blood cultures were initiated with RPMI-1640 (80%) supplemented with FBS (20%), stimulated with mitogen and incubated at 37°C for 44h. At the 44th hour, cytochalasin-B (6µg/mL) was added, and the cultures were incubated for 28h more. The cells were harvested with a pre-chilled hypotonic solution (0.075M) and fixed with a Carnoy's solution (methanol/acetic acid 5:1). Giemsa- and propidium-iodide-stained cells affixed to slides for microscopy were scored manually and automatically with the micronucleus scoring software from MetaSystems. The micronucleus frequencies determined in the Giemsa-stained cells by manual and automated scoring were 23.6% different (P<0.0001) with an efficiency of 24.9%. Slides stained with propidium iodide are a better choice for automated scoring than Giemsa-stained ones.
对受电离辐射个体的外周血淋巴细胞中的微核进行评分是一种快速生物剂量测定法。五名个体的外周血淋巴细胞在体外以0.76Gy/分钟的剂量率暴露于0 - 5Gy的钴-60γ辐射。血液培养用补充有20%胎牛血清的80% RPMI - 1640培养基起始，用促有丝分裂原刺激，并在37°C下孵育44小时。在第44小时，加入细胞松弛素B（6μg/mL），培养物再孵育28小时。用预冷的低渗溶液（0.075M）收获细胞，并用卡诺氏溶液（甲醇/乙酸5:1）固定。固定在载玻片上用于显微镜检查的吉姆萨染色和碘化丙啶染色的细胞，使用MetaSystems的微核评分软件进行手动和自动评分。通过手动和自动评分在吉姆萨染色细胞中确定的微核频率相差23.6%（P<0.0001），效率为24.9%。碘化丙啶染色的载玻片比吉姆萨染色的载玻片更适合自动评分。
