Abstract
This report describes a feasibility study concerning the use of a visible diode laser for two important fluorescence applications in a flow cytometer. With a 3 mW 635 nm. diode laser, we performed immunofluorescence measurements using the fluorophore allophycocyanin (APC). We have measured CD8 positive lymphocytes with a two-step labeling procedure and the resulting histograms showed good separation between the negative cells and the dim and the bright fluorescent subpopulations. As a second fluorescence application, we chose DNA analysis with the recently developed DNA/ RNA stains TOTO-3 and TO-PRO-3. In our setup TO-PRO-3 yielded the best results with a CV of 3.4%. Our results indicate that a few milliwatts of 635 nm light from a visible diode laser is sufficient to do single color immunofluorescence measurements with allophycocyanin and DNA analysis with TO-PRO-3. The major advantages of using a diode laser in a flow cytometer are the small size, the low price, the high efficiency, and the long lifetime.
Original language | Undefined |
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Pages (from-to) | 267-271 |
Number of pages | 5 |
Journal | Cytometry |
Volume | 15 |
Issue number | 3 |
DOIs | |
Publication status | Published - 1994 |
Keywords
- TO-PRO-3
- TOTO-3
- IR-60722
- CD8
- APC
- METIS-129465
- Miniaturization
- Flow cytometry
- Instrumentation
- allophycocyanin
- DNA/RNA staining