Multiple wavelength illumination in flow cytometry using a single arc lamp and a dispersing element

B.G. de Grooth, M. van Dam, N.C. Swart, A. Willemsen, Jan Greve

    Research output: Contribution to journalArticleAcademic

    6 Citations (Scopus)
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    Abstract

    The principle of a multiple wavelength illumination method for flow cytometers, based upon a combination of a helium-neon laser and an arc lamp as illumination sources is described. By using a prism, the light from the arc lamp is dispersed and the different colors are imaged at different places on the sample stream. The small angle light scattering from the helium-neon laser light is measured as a relevant parameter and serves as a trigger signal for subsequent measurements of fluorescence or scattering of light from the arc lamp. Two experimental systems are described utilizing this principle: a system where the emission is detected orthogonally with respect to the direction of the illumination beams, and an epi-illumination system. With the orthogonal set-up multiple wavelength right angle scattering measurements are possible. This is illustrated by showing that the orthogonal scattering from erythrocytes is strongly dependent on the illumination wavelength. It is further shown that the apparatus is suitable for the measurement of intracellular pH using the pH dependence of the excitation spectrum of fluorescein. The epi-illumination system allows excitation of two (or more) fluorescent dyes with different excitation spectra. In this case the emission spectra of the fluorescent dyes may overlap substantially. This is shown by simultaneous measurement of DNA and protein of Chinese hamster lung cells using mitramycin and tetramethyl rhodamin isothiocyanate (TRITC).
    Original languageUndefined
    Pages (from-to)445-452
    JournalCytometry
    Volume8
    Issue number5
    DOIs
    Publication statusPublished - 1987

    Keywords

    • IR-60749
    • DNA-protein staining
    • prism
    • Intracellular pH measurement

    Cite this

    de Grooth, B. G., van Dam, M., Swart, N. C., Willemsen, A., & Greve, J. (1987). Multiple wavelength illumination in flow cytometry using a single arc lamp and a dispersing element. Cytometry, 8(5), 445-452. https://doi.org/10.1002/cyto.990080503
    de Grooth, B.G. ; van Dam, M. ; Swart, N.C. ; Willemsen, A. ; Greve, Jan. / Multiple wavelength illumination in flow cytometry using a single arc lamp and a dispersing element. In: Cytometry. 1987 ; Vol. 8, No. 5. pp. 445-452.
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    abstract = "The principle of a multiple wavelength illumination method for flow cytometers, based upon a combination of a helium-neon laser and an arc lamp as illumination sources is described. By using a prism, the light from the arc lamp is dispersed and the different colors are imaged at different places on the sample stream. The small angle light scattering from the helium-neon laser light is measured as a relevant parameter and serves as a trigger signal for subsequent measurements of fluorescence or scattering of light from the arc lamp. Two experimental systems are described utilizing this principle: a system where the emission is detected orthogonally with respect to the direction of the illumination beams, and an epi-illumination system. With the orthogonal set-up multiple wavelength right angle scattering measurements are possible. This is illustrated by showing that the orthogonal scattering from erythrocytes is strongly dependent on the illumination wavelength. It is further shown that the apparatus is suitable for the measurement of intracellular pH using the pH dependence of the excitation spectrum of fluorescein. The epi-illumination system allows excitation of two (or more) fluorescent dyes with different excitation spectra. In this case the emission spectra of the fluorescent dyes may overlap substantially. This is shown by simultaneous measurement of DNA and protein of Chinese hamster lung cells using mitramycin and tetramethyl rhodamin isothiocyanate (TRITC).",
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    year = "1987",
    doi = "10.1002/cyto.990080503",
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    de Grooth, BG, van Dam, M, Swart, NC, Willemsen, A & Greve, J 1987, 'Multiple wavelength illumination in flow cytometry using a single arc lamp and a dispersing element' Cytometry, vol. 8, no. 5, pp. 445-452. https://doi.org/10.1002/cyto.990080503

    Multiple wavelength illumination in flow cytometry using a single arc lamp and a dispersing element. / de Grooth, B.G.; van Dam, M.; Swart, N.C.; Willemsen, A.; Greve, Jan.

    In: Cytometry, Vol. 8, No. 5, 1987, p. 445-452.

    Research output: Contribution to journalArticleAcademic

    TY - JOUR

    T1 - Multiple wavelength illumination in flow cytometry using a single arc lamp and a dispersing element

    AU - de Grooth, B.G.

    AU - van Dam, M.

    AU - Swart, N.C.

    AU - Willemsen, A.

    AU - Greve, Jan

    PY - 1987

    Y1 - 1987

    N2 - The principle of a multiple wavelength illumination method for flow cytometers, based upon a combination of a helium-neon laser and an arc lamp as illumination sources is described. By using a prism, the light from the arc lamp is dispersed and the different colors are imaged at different places on the sample stream. The small angle light scattering from the helium-neon laser light is measured as a relevant parameter and serves as a trigger signal for subsequent measurements of fluorescence or scattering of light from the arc lamp. Two experimental systems are described utilizing this principle: a system where the emission is detected orthogonally with respect to the direction of the illumination beams, and an epi-illumination system. With the orthogonal set-up multiple wavelength right angle scattering measurements are possible. This is illustrated by showing that the orthogonal scattering from erythrocytes is strongly dependent on the illumination wavelength. It is further shown that the apparatus is suitable for the measurement of intracellular pH using the pH dependence of the excitation spectrum of fluorescein. The epi-illumination system allows excitation of two (or more) fluorescent dyes with different excitation spectra. In this case the emission spectra of the fluorescent dyes may overlap substantially. This is shown by simultaneous measurement of DNA and protein of Chinese hamster lung cells using mitramycin and tetramethyl rhodamin isothiocyanate (TRITC).

    AB - The principle of a multiple wavelength illumination method for flow cytometers, based upon a combination of a helium-neon laser and an arc lamp as illumination sources is described. By using a prism, the light from the arc lamp is dispersed and the different colors are imaged at different places on the sample stream. The small angle light scattering from the helium-neon laser light is measured as a relevant parameter and serves as a trigger signal for subsequent measurements of fluorescence or scattering of light from the arc lamp. Two experimental systems are described utilizing this principle: a system where the emission is detected orthogonally with respect to the direction of the illumination beams, and an epi-illumination system. With the orthogonal set-up multiple wavelength right angle scattering measurements are possible. This is illustrated by showing that the orthogonal scattering from erythrocytes is strongly dependent on the illumination wavelength. It is further shown that the apparatus is suitable for the measurement of intracellular pH using the pH dependence of the excitation spectrum of fluorescein. The epi-illumination system allows excitation of two (or more) fluorescent dyes with different excitation spectra. In this case the emission spectra of the fluorescent dyes may overlap substantially. This is shown by simultaneous measurement of DNA and protein of Chinese hamster lung cells using mitramycin and tetramethyl rhodamin isothiocyanate (TRITC).

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    KW - DNA-protein staining

    KW - prism

    KW - Intracellular pH measurement

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    JF - Cytometry

    SN - 0196-4763

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