Evaluation of Excitation Light Sources for Incident Immunofluorescence Microscopy

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Published inApplied Microbiology Vol. 30; no. 4; pp. 616 - 624
Main Authors Thomson, L A, Hageage, G J
Format Journal Article
LanguageEnglish
Published United States American Society for Microbiology 01.10.1975
Subjects
Evaluation Studies as Topic
Fluorescent Antibody Technique
Halogens
Light
Mercury
Methods
Microscopy, Fluorescence - instrumentation
Xenon
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A variety of fluorescent excitation light sources were compared using a standard fluorescein solution or a bacterial conjugate with immunofluorescent microscopy. Quantitative data were obtained with microscope photometric apparatus. Both the quantitative data and comparative conjugate titering suggest that the 450-W xenon arc excited significantly more fluorescence than did the more commonly used 250-W mercury arc or the 100-W halogen lamp. The conjugate could be diluted 4 to 32 times more using the 450-W xenon. Additional advantages of 450-W xenon excitation include sufficient energy of wavelengths between 470 to 490 mm, thus permitting narrow-band excitation resulting in less autofluorescence and the ability to perform fluorescent-antibody procedures without the darkening of ambient room light.
A variety of fluorescent excitation light sources were compared using a standard fluorescein solution or a bacterial conjugate with immunofluorescent microscopy. Quantitative data were obtained with microscope photometric apparatus. Both the quantitative data and comparative conjugate titering suggest that the 450-W xenon arc excited significantly more fluorescence than did the more commonly used 250-W mercury arc or the 100-W halogen lamp. The conjugate could be diluted 4 to 32 times more using the 450-W xenon. Additional advantages of 450-W xenon excitation include sufficient energy of wave lengths between 470 to 490 mm, thus permitting narrow-band excitation resulting in less autofluorescence and the ability to perform fluorescent-antibody procedures without the darkening of ambient room light.
Author L. A. Thomson
G. J. Hageage
AuthorAffiliation National Caries Program and Laboratory of Microbiology and Immunology, National Institute of Dental Research, Bethesda, Maryland 20014
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Present address: Pathology Department, St. Vincent Hospital, Toledo, Ohio 43608.
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4117182 - Eur J Immunol. 1972 Feb;2(1):1-4
4563708 - Clin Exp Immunol. 1972 Aug;11(4):483-94
4945674 - Ann N Y Acad Sci. 1971 Jun 21;177:414-29
4882659 - Z Wiss Mikrosk. 1967 Nov;68(3):129-42
4119491 - Histochemie. 1973;33(1):61-70
4929952 - Histochemie. 1971;25(4):329-43
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SubjectTerms Evaluation Studies as Topic
Fluorescent Antibody Technique
Halogens
Light
Mercury
Methods
Microscopy, Fluorescence - instrumentation
Xenon
Title Evaluation of Excitation Light Sources for Incident Immunofluorescence Microscopy
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Volume 30
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