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Separation of mouse epidermal basal and differentiating cells for microflow fluorometric measurements: a methodologic
Virchows Archiv. B, Cell Pathology
|May 26, 1976
Summary
Microflow fluorometry (MFF) offers improved DNA content resolution for epidermal basal cells and lymphocytes. Pepsin treatment enhances lymphocyte fluorescence, enabling accurate DNA measurements for cell cycle analysis.
Area of Science:
- Cell Biology
- Biophysics
- Dermatology
Background:
- Accurate DNA content measurement is crucial for understanding cell proliferation.
- Conventional methods like Feulgen microspectrophotometry have limitations in resolution.
- Microflow fluorometry (MFF) offers a potential advancement in cytophotometry.
Purpose of the Study:
- To compare DNA content measurement in lymphocytes and epidermal basal cells using MFF and conventional methods.
- To evaluate the effect of pepsin treatment on lymphocyte DNA measurements.
- To determine the cell cycle distribution of epidermal basal cells.
Main Methods:
- Microflow fluorometry (MFF) for DNA content analysis.
- Combined mechanical and enzymatic method for epidermal basal cell isolation.
- Feulgen microspectrophotometry on histologic sections.
- Pepsin treatment of lymphocytes prior to MFF.
Main Results:
- MFF demonstrated improved resolution compared to Feulgen microspectrophotometry.
- Pepsin treatment of lymphocytes increased their fluorescence intensity to match basal cells.
- Cell cycle analysis of basal cells showed changes in S phase correlating with 3H-Tdr labeling index.
Conclusions:
- MFF provides superior resolution for DNA content analysis of epidermal cells and lymphocytes.
- Pepsin treatment is advantageous for accurate MFF-based DNA measurements.
- MFF facilitates reliable cell cycle distribution determination in epidermal basal cells.