FLOW CYTOMETRIC MEASUREMENT OF TOTAL DNA CONTENT AND INCORPORATED BROMODEOXYURIDINE
FLOW CYTOMETRIC MEASUREMENT OF TOTAL DNA CONTENT AND INCORPORATED BROMODEOXYURIDINE
复制标题
DOI：
10.1073/pnas.80.18.5573
复制
发表时间：
1983-01-01
期刊：
影响因子：
--
通讯作者：
GRAY, JW
中科院分区：
文献类型：
--
作者：
DOLBEARE, F;GRATZNER, H;GRAY, JW
开发了一种程序，用于同时流式细胞术测量细胞DNA含量和掺入细胞DNA中的BrdUrd [溴脱氧尿苷]量。使用碘化丙啶作为总细胞DNA的荧光探针，使用抗BrdUrd的单克隆抗体作为BrdUrd掺入DNA的探针。使用荧光素标记的山羊抗小鼠抗体来荧光标记结合的抗BrdUrd探针。用BrdUrd标记30分钟的中国仓鼠卵巢细胞测量的双变量DNA/BrdUrd分布清楚地显示G1和G2 M期细胞具有低BrdUrd相关荧光，S期细胞具有高BrdUrd相关荧光。根据BrdUrd脉冲标记后定期采集的样本测量的双变量分布，估计中国仓鼠卵巢细胞的细胞周期穿越率。还将双变量DNA/BrdUrd分布应用于C3 H鼠骨髓细胞对用1-β-BrdUrd处理的体内反应的分析中。D-阿拉伯呋喃糖基胞嘧啶（araC）。在用araC处理后的不同时间，测量取自用BrdUrd脉冲标记的小鼠的骨髓细胞的双变量分布。所得的DNA/BrdUrd序列显示了从araC中回收的动力学，并允许区分araC灭菌的细胞。
A procedure was developed for simultaneous flow cytometric measurement of cellular DNA content and amount of BrdUrd [bromodeoxyuridine] incorporated into cellular DNA. Propidium iodide was used as a fluorescent probe for total cellular DNA and a monoclonal antibody against BrdUrd was used as a probe for BrdUrd incorporated into DNA. Fluorescein-labeled goat anti-mouse antibody was used to fluorescently label the bound anti-BrdUrd probe. Bivariate DNA/BrdUrd distributions measured for Chinese hamster ovary cells labeled for 30 min with BrdUrd clearly show the G1- and G2M-phase cells to have low BrdUrd-linked fluorescence and the S-phase cells to have high BrdUrd-linked fluorescence. Cell cycle traverse rates were estimated for Chinese hamster ovary cells from bivariate distributions measured for samples taken periodically after pulse labeling with BrdUrd. Bivariate DNA/BrdUrd distributions were also applied in the analysis of the response of C3H murine bone marrow cells to treatment in vivo with 1-.beta.-D-arabinofuranosylcytosine (araC). Bivariate distributions were measured for bone marrow cells taken from mice that were pulse labeled with BrdUrd at various times after treatment with araC. The resulting DNA/BrdUrd sequences show the kinetics of recovery from araC and allow discrimination of the araC sterilized cells.