A METHOD TO MEASURE THE DURATION OF DNA-SYNTHESIS AND THE POTENTIAL DOUBLING TIME FROM A SINGLE SAMPLE
A METHOD TO MEASURE THE DURATION OF DNA-SYNTHESIS AND THE POTENTIAL DOUBLING TIME FROM A SINGLE SAMPLE
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DOI：
10.1002/cyto.990060618
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发表时间：
1985-01-01
期刊：
影响因子：
--
通讯作者：
KARCHER, H
中科院分区：
文献类型：
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作者：
BEGG, AC;MCNALLY, NJ;KARCHER, H
本文描述了一种方法，利用溴脱氧尿苷（BrdUrd）标记后数小时采集的单个细胞样本的数据，可以计算DNA合成时间Ts。该方法包括使用BrdUrd掺入（绿色荧光，fitc标记的抗BrdUrd-DNA抗体）和总DNA含量（红色荧光，碘化丙啶）的流式细胞术数据进行简单计算。brdurd标记的细胞在S期的运动可以通过测量其相对于G1和G2细胞的平均红色荧光来量化。假设标记细胞向G2方向的运动随时间呈线性，则可以通过测量它们在任意时刻的相对运动来计算t。该方法在体外细胞和[小鼠]骨髓及肿瘤细胞上进行了实验。这些组织的t值与已发表的估计值基本一致。
A method is described whereby the DNA synthesis time, Ts, can be calculated using data of a single sample of cells taken several hours after labelling with bromodeoxyuridine (BrdUrd). The method involves a simple calculation using flow cytometry data of BrdUrd incorporation (green fluorescence, FITC-labelled anti-BrdUrd-DNA antibody) and total DNA content (red fluorescence, propidium iodide). The movement of BrdUrd-labelled cells through the S phase can be quantified by measuring their mean red fluorescence relative to that of G1 and G2 cells. Assuming the movement of the labelled cells toward G2 is linear with time, Ts can be calculated by measuring their relative movement at any one time. The method was tested on cells in vitro and on [mouse] bone marrow and tumor cells in vivo. Reasonable agreement was seen with published estimates of Ts for these tissues.