Abstract
Detection of proliferating cells based on bromodeoxyuridine (BrdU) incorporation and determination of phenotype by immunofluorescence labeling are standard approaches for studying stem and progenitor cell populations in developing and adult tissue as well as in histopathology studies. We describe incorporation of different halogenated thymidine analogs for temporal discrimination of cell cycle in the rat. With equimolar delivery, these analogs are suitable for quantitative histological studies including assessment of the regulation of proliferation, clonal analysis and simultaneous profiling of cell phenotype relative to proliferative history.
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Acknowledgements
This work was supported by National Institutes of Health AG20047.
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Supplementary information
Supplementary Fig. 1
Immunohistochemical detection can discriminate between CldU and IdU. (PDF 5294 kb)
Supplementary Fig. 2
Immunohistochemistry for CldU and IdU detects an equivalent population of proliferating cells, but only with equimolar delivery of the halogenated thymidine analogs. (PDF 4767 kb)
Supplementary Fig. 3
Temporal separation of CldU and IdU administration permits the determination of neural stem/progenitor cell proliferative history. (PDF 3317 kb)
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Vega, C., Peterson, D. Stem cell proliferative history in tissue revealed by temporal halogenated thymidine analog discrimination. Nat Methods 2, 167–169 (2005). https://doi.org/10.1038/nmeth741
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DOI: https://doi.org/10.1038/nmeth741
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