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      A chemical method for fast and sensitive detection of DNA synthesis in vivo.

      Proceedings of the National Academy of Sciences of the United States of America

      Animals, Time Factors, Sensitivity and Specificity, Reproducibility of Results, NIH 3T3 Cells, Molecular Structure, Mice, chemistry, analysis, analogs & derivatives, Deoxyuridine, DNA Replication, biosynthesis, DNA

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          Abstract

          We have developed a method to detect DNA synthesis in proliferating cells, based on the incorporation of 5-ethynyl-2'-deoxyuridine (EdU) and its subsequent detection by a fluorescent azide through a Cu(I)-catalyzed [3 + 2] cycloaddition reaction ("click" chemistry). Detection of the EdU label is highly sensitive and can be accomplished in minutes. The small size of the fluorescent azides used for detection results in a high degree of specimen penetration, allowing the staining of whole-mount preparations of large tissue and organ explants. In contrast to BrdU, the method does not require sample fixation or DNA denaturation and permits good structural preservation. We demonstrate the use of the method in cultured cells and in the intestine and brain of whole animals.

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          Author and article information

          Journal
          18272492
          10.1073/pnas.0712168105
          2268151

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