Flow microfluorometric analysis of nuclear DNA in cells from solid tumors and cell suspensions
Flow microfluorometric analysis of nuclear DNA in cells from solid tumors and cell suspensions
复制标题
对实体瘤细胞和细胞悬液中的核 DNA 进行流式显微荧光分析
DOI：
10.1007/bf02889282
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发表时间：
1977
期刊：
影响因子：
--
通讯作者：
Lars L. Vindeløv
中科院分区：
文献类型：
--
作者：
Lars L. Vindeløv
本文介绍了一种用于流动显微荧光DNA分析的一步法制备细胞核的方法，用非离子洗涤剂Nonidet P40裂解细胞膜。单细胞悬液和细针活检获得的固体组织标本可以同样好地制备，固体组织细胞的核被分开释放。在含有溴化乙锭或碘化丙啶的染色溶液中进行裂解。由于荧光素与RNA结合而产生的荧光可被RNA-Se瞬时消除，而与DNA二级结合部位的荧光素结合则被氯化钠抑制。制备时间为10min，样品至少稳定12h。使用基本版本的方法，可用于所有受测细胞类型的结果，包括白细胞、骨髓、肝细胞、人淋巴瘤、人乳腺癌和人肺癌、JB-1腹水肿瘤。描述了由此产生的两种改进的技术。用溴化乙锭和碘化丙啶的类似物染色白细胞的不同。
SummaryA one-step procedure for the preparation of nuclei for flow micro fluorometric DNA analysis is described.The membranes of the cells were lysed by the non-ionic detergent Nonidet P40. Single-cell suspensions, and specimens of solid tissues obtained with fine-needle biopsy, could be prepared equally well as the nuclei of solid tissue cells were released separately. Lysis was performed in the staining solution containing either ethidium bromide or propidium iodide. Fluorescence due to fluorochrome binding to RNA, was abolished instantaneously by the presence of RNA-se, and fluorochrome binding to secondary binding sites in DNA was inhibited with NaCl. The preparation time was 10 min and the samples were stable for a minimum of 12 h.With the basic version of the method, usable, but not always optimal, results were obtained in all the cell types tested: four different mouse ascites tumors, leucocytes, bone-marrow, liver cells, human lymphomas, human carcinomas of the breast and lung, mouse mammary carcinoma and solid JB-1 tumor.The method was further optimized for the JB-1 ascites tumour. The resulting two modified techniques are described.Differences in the staining of leucocytes with the analogues ethidium bromide and propidium iodide were demonstrated.