8. In Vivo Labeling and Analysis of Epidermal Stem Cells
| Abstract |
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To identify slowly cycling (label-retaining) keratinocytes in the cutaneous epithelium of mice, newborn female pups were injected
with [3H]-thymidine twice daily for 3 d beginning the third day after birth. At 8–10 wk of age, skin from the dorsal side of the
mice was removed and fixed. The skin was processed for paraffin embedding, and serial sections were prepared. Dewaxed and
ethanol-treated slides were dried and dipped into autoradiographic emulsion and allowed to dry in light tight box for 4 wk
at 4°C. After fixation with photographic fixer, the slides were washed, lightly stained, and viewed with a light microscope
to observe and quantify cells that retained the labeling. Label-retaining cells were identified by the black silver grains
over their nuclei.
Affiliation(s): (2) Department of Dermatology, Columbia University College of Physicians and Surgeons, New York, NY
Book Title: Epidermal Cells: Methods and Protocols
Series: Methods in Molecular Biology | Volume: 289 | Pub. Date: Oct-20-2004 | Page Range: 73-78 | DOI: 10.1385/1-59259-830-7:073
Subject: Cell Biology
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