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Selection of viable cells with known DNA content
Cytometry
|September 1, 1980
Summary
This study shows that Hoechst 33342 and fluorescein energy transfer allows cell sorting based on DNA content and viability using a single UV laser. This method enables precise selection of cell populations with specific DNA and viability characteristics.
Area of Science:
- Cell Biology
- Biophysics
- Flow Cytometry
Background:
- Flow cytometry is crucial for analyzing cell populations.
- Distinguishing cell viability and DNA content simultaneously is challenging.
- Hoechst 33342 and fluorescein diacetate are common cellular stains.
Purpose of the Study:
- To demonstrate resonance energy transfer between Hoechst 33342 and fluorescein.
- To establish a method for selecting cells based on DNA content and viability using a single UV laser.
- To confirm the accessibility of fluorescein to the nucleus.
Main Methods:
- Cells were doubly stained with Hoechst 33342 (DNA content) and fluorescein diacetate (viability).
- A single UV laser flow sorter was utilized for cell selection.
- Static fluorescence microscopy and a dual laser flow sorter were employed for verification.
Main Results:
- Resonance energy transfer was confirmed between Hoechst 33342 and fluorescein in doubly stained cells.
- Sensitized fluorescein emission, combined with DNA emission, enabled selection of cell populations.
- Cells with known DNA content and viability were successfully sorted.
- Fluorescein liberated by esterases demonstrated free accessibility to the nucleus.
Conclusions:
- Single UV laser flow sorting is effective for selecting cells based on DNA content and viability.
- Resonance energy transfer is a viable mechanism for multiplexed cellular analysis.
- The accessibility of intracellular fluorescein to the nucleus is confirmed.