Abstract
Labelling and identification of proliferating cells is important for the study of physiological or pathological processes in high-content screening (HCS) assays. Here we describe ethynyl deoxyuridine (EdU) as a biomarker for the assessment of cell proliferation and clearly demonstrate the feasibility of the EdU-labelling method for use in HCS assays. EdU detection is highly robust, reproducible, technically simple, and well suited for automated segmentation, which provides an excellent alternative for setting up multiplexed HCS assays of siRNA, miRNA and small-molecule libraries.
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Hanev SA, La Pan P, Pan J, Zhang J. High-content screening moves to the front of the line. Drug Discov Today, 2006, 11(19–20): 889–894
Salic A, Mitchison TJ. A chemical method for fast and sensitive detection of DNA synthesis in vivo. Proc Natl Acad Sci USA, 2008, 105: 2415–2420
Diermeier-Daucher S, Clarke ST, Hill D, Vollmann-Zwerenz A, Bradford JA, Brockhoof G. Cell type specific applicability of 5-ethynyl-2′-deoxyuridine (EdU) for dynamic proliferation assessment in flow cytometry. Cytometry A, 2009, 75(6): 535–546
Meneni S, Ott I, Sergeant CD, Sniady A, Gust R, Dembinski R. 5-Alkynyl-2′-deoxyuridines: chromatography-free synthesis and cytotoxicity evaluation against human breast cancer cells. Bioorg Med Chem, 2007, 15 (8): 3082–3088
Mujumdar RB, Ernst LA, Mujumdar SR, Lewis CJ, Waggoner AS. Cyanine dye labeling reagents: Sulfoindocyanine succinimidyl esters. Bioconjug Chem, 1993, 4(2):105–111
De Boer RJ, Mohri H, Ho DD, Perelson AS. Estimating average cellular turnover from 5-bromo-2′-deoxyuridine (BrdU) measurements. Proc R Soc B, 2003, 270(1517): 849–858
von Trotha JW, Egger B, Brand AH. Cell proliferation in the drosophila adult brain revealed by clonal analysis and bromodeoxyuridine labelling. Neural Dev, 2009, 4, 9
Rothaeusler K, Baumgarth N. Evaluation of intranuclear BrdU detection procedures for use in multicolor flow cytometry. Cytometry A, 2006, 69(4): 249–259
Gudla PR, Nandy K, Collins J, Meaburn KJ, Misteli T, Lockett SJ. A high-throughput system for segmenting nuclei using multiscale techniques. Cytometry A, 2008, 73(5): 451–466
Yan P, Zhou X, Shah M, Wong ST. Automatic segmentation of high-throughput RNAi fluorescent cellular images. IEEE Trans Inf Technol Biomed, 2008, 12(1): 109–117
Havden MS, Ghosh S. Signaling to NF-κB. Genes Dev, 2004, 18: 2195–2224
Havden MS, Ghosh S. Shared principles in NF-kappaB signaling. Cell, 2008, 132(3): 344–362
Fiedler MA, Wernke-Dollries K, Stark JM. Inhibition of TNF-α-induced NF-κB activation and IL-8 release in A549 cells with the proteasome inhibitor MG-132. Am J Respir Cell Mol Biol, 1998, 19: 259–268
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Dedicated to Prof. Ronald Breslow on the occasion of his 80th birthday
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Chen, M., Qu, D., Chi, W. et al. 5-Ethynyl-2′-deoxyuridine as a molecular probe of cell proliferation for high-content siRNA screening assay by “click” chemistry. Sci. China Chem. 54, 1702–1710 (2011). https://doi.org/10.1007/s11426-011-4398-4
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DOI: https://doi.org/10.1007/s11426-011-4398-4