Visible Diode Lasers can be Used for Flow Cytometric Immunofluorescence and DNA Analysis

Richard M.P. Doornbos, R.M.P. Doornbos, B.G. de Grooth, Yvonne M. Kraan, Carel J. van der Poel, Jan Greve

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    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 languageUndefined
    Pages (from-to)267-271
    Number of pages5
    JournalCytometry
    Volume15
    Issue number3
    DOIs
    Publication statusPublished - 1994

    Keywords

    • TO-PRO-3
    • TOTO-3
    • IR-60722
    • CD8
    • APC
    • METIS-129465
    • Miniaturization
    • Flow cytometry
    • Instrumentation
    • allophycocyanin
    • DNA/RNA staining

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