| 许雅苹,郑晓辉,房纯正,翁乐斌,黄黎月.DAPI染色法在流式细胞术中去除死细胞影响的探讨[J].,2019,19(9):1660-1664 |
| DAPI染色法在流式细胞术中去除死细胞影响的探讨 |
| A Methodology Study on Elimination the Influence of Dead Cells Stained with DAPI |
| 投稿时间：2018-10-31 修订日期：2018-11-27 |
| DOI：10.13241/j.cnki.pmb.2019.09.012 |
| 中文关键词: DAPI 流式细胞术 死细胞 |
| 英文关键词: DAPI Flow cytometry Dead cells |
| 基金项目:国家自然科学基金项目（81602500）；福建省卫计委中青年项目（2016-2-80）；2017年福建省高校杰出青年科研人才培育计划项目；博士科研启动经费自助（K2016-07） |
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| 中文摘要: |
| 摘要 目的：探索利用DAPI染色法在流式细胞检测过程中去除死细胞，消除死细胞对流式结果的影响。方法：建立原位肝癌小鼠模型，流式检测其中死细胞对脾脏中骨髓来源抑制性细胞（myeloid-derived suppressor cell，MDSC）结果的影响，并以碘化丙啶（propidiumiodide，PI）为对照，探索不同DAPI浓度、不同染色时间对死细胞比例的影响。结果：流式细胞术中去除死细胞影响时DAPI染色浓度低于1 g/mL时，其染色结果影响不大，浓度大于1 g/mL，死细胞比例明显升高；当DAPI使用浓度为0.2 g/mL时，在30钟内死细胞染色结果与PI相比差别无统计学差异，当染色时间大于30 min时，死细胞的比例显著升高。结论：DAPI染色法应用于流式细胞术中去除死细胞是一种可靠的检测方法，且简单易行，推荐使用方法为0.2 g/mL，染色时间1-5 min。 |
| 英文摘要: |
| ABSTRACT Objective: To explore the method of DAPI staining in the process of flow cytometry to remove dead cells and elimi- nate the influence of dead cells on flow cytometry results. Methods: An orthotopic transplantion tumor model of hepatoma in mice was used to detect the dead cells effect on the result of MDSC in splencyte by flow cytometry. PI was used as a positive control to contrast the influence of different DAPI concentration and staining time on the proportion of dead cells. Results: The data showed that while the DAPI concentration was less than 1 g/mL, there had a similar result of the proportion of dead cells compared with PI staining; however, if the concentration of DAPI was more than 1 g/mL, the proportion of dead cells was increased. Stained the cells with 0.2 g/mL of DAPI within 30 minutes had a little influence on the proportion of dead cells. But when the staining time exceed 30 minutes, the proportion of dead cells increased obviously. Conclusion: The DAPI staining method is a reliable and easy detection method applied to remove dead cells effect on the flow cytometry, 0.2 g/mL of DAPI stained 1-5 minutes was recommend. |
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