Abstract
DNA labeling in vivo using nucleoside analogues is a current experimental approach to determine cell proliferation rates in cell cultures and tissues. It has also been successfully used to localize adult stem cell niches through the identification of nucleoside label-retaining cells (LRC) in long-term experiments. A major hindrance of this methodology relies on the selection of adequate procedures to quantify the nucleoside analogue content from image data files. Here we propose a simple procedure using Fiji image processing software to accurately calculate nucleoside analogue retaining chromatin/total chromatin (LRC/DAPI) signal ratios in the well-known mouse hair follicle stem cell niche.
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Acknowledgements
This work was supported by grants of the Spanish Ministerio de Economía y Competitividad (SAF 11-23493) and the Comunidad Autónoma de Madrid (SkinModel, CAM S10/BMD-2359) to J.E.; E.C and M.I.C. are supported by Ph.D. fellowship grants of the Spanish Ministerio de Educación and Universidad Autónoma de Madrid, respectively.
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Carrasco, E., Calvo, M.I., Espada, J. (2014). DNA Labeling In Vivo: Quantification of Epidermal Stem Cell Chromatin Content in Whole Mouse Hair Follicles Using Fiji Image Processing Software. In: Stockert, J., Espada, J., Blázquez-Castro, A. (eds) Functional Analysis of DNA and Chromatin. Methods in Molecular Biology, vol 1094. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-706-8_7
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DOI: https://doi.org/10.1007/978-1-62703-706-8_7
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