Carbajo S, Orfao A, Alberca V, Ciudad J, López A, Hernández L C, Carbajo-Pérez E
Departamento de Anatomía e Histología Humansas, Universidad de Salamanca, Spain.
Anal Cell Pathol. 1995 Jan;8(1):15-25.
Tissue sections and single cell suspensions of rat thymus were analyzed by flow and image cytometry to study the effects of different doses of bromodeoxyuridine (BrdU) (10, 50 and 100 mg/kg and different labelling periods (15, 30 and 60 min) on the BrdU-labelling index. The proportions of BrdU-labelled cells, as analyzed by flow cytometry, were similar in the different groups of our study. However, the quality of the results was not exactly the same, since when the BrdU dose decreased, the separation between BrdU-positive S-phase cells and both the G0/G1 and the G2/M BrdU-negative cells on a DNA/BrdU histogram became less clear. On the other hand, the proportion of BrdU-labelled cells in tissue sections was greater in the groups of animals that received doses of 100 mg/kg and were killed 15, 30 or 60 min later than in the sections obtained from the remaining groups. Our results show that both dose and exposure time to BrdU may influence the final results when cell proliferation is assessed, the variations obtained clearly depending on the technique used for the immunological detection of BrdU-positive S-phase cells.
采用流式细胞术和图像细胞术分析大鼠胸腺组织切片和单细胞悬液，以研究不同剂量的溴脱氧尿苷（BrdU）（10、50和100mg/kg）以及不同标记时间（15、30和60分钟）对BrdU标记指数的影响。在我们的研究中，通过流式细胞术分析，不同组中BrdU标记细胞的比例相似。然而，结果的质量并不完全相同，因为当BrdU剂量降低时，在DNA/BrdU直方图上，BrdU阳性S期细胞与G0/G1期和G2/M期BrdU阴性细胞之间的分离变得不那么清晰。另一方面，在接受100mg/kg剂量并在15、30或60分钟后处死的动物组的组织切片中，BrdU标记细胞的比例高于从其余组获得的切片。我们的结果表明，在评估细胞增殖时，BrdU的剂量和暴露时间都可能影响最终结果，所获得的变化显然取决于用于免疫检测BrdU阳性S期细胞的技术。
