Detection of cell-surface antigens using antibody-conjugated fluorospheres (ACF) : application for six-color immunofluorescence
Antibody-conjugated fluorospheres (ACF) were used to phenotype murine leukocytes by flow cytometric analysis. Multicolor immunofluoresence (beyond simultaneous 4-color analysis) is limited by the availability of specific antibody-fluorochrome chrome conjugates and even further restricted by the spec...
| Publié dans: | BioTechniques. - 1991. - 21(1996), 3 vom: 15. Sept., Seite 498-503 |
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| Format: | Article |
| Langue: | English |
| Publié: |
1996
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| Accès à la collection: | BioTechniques |
| Sujets: | Comparative Study Journal Article Antibodies, Monoclonal Antigens, Differentiation, T-Lymphocyte Antigens, Ly Antigens, Surface Fluorescent Dyes Histocompatibility Antigens Class II Macrophage-1 Antigen Organometallic Compounds plus... |
| Résumé: | Antibody-conjugated fluorospheres (ACF) were used to phenotype murine leukocytes by flow cytometric analysis. Multicolor immunofluoresence (beyond simultaneous 4-color analysis) is limited by the availability of specific antibody-fluorochrome chrome conjugates and even further restricted by the spectral emission overlap of many of the fluorochromes when used in combination. Fluorospheres possessing unique excitation/emission spectra can provide much needed versatility to existing protocols of multicolor fluorescence. SKY BLUE (647 nm excitation, 730 nm emission) fluorospheres conjugated to CD11b monoclonal antibody were used in combination with the monoclonal antibodies IAd-FITC, L3T4 (CD4)-PE, LYT2 (CD8)-APC, THY1.2 (CD90)-biotin and B220 (CD45R)-RED613 for the simultaneous detection of six distinct cell-surface antigens in a mixed cell population. All fluorescence signals were resolved, and comparison of results from five-, six- and single-color samples indicated that the percentages of cells positive for specific surface antigens were equivalent |
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| Description: | Date Completed 11.02.1997 Date Revised 28.09.2018 published: Print Citation Status MEDLINE |
| ISSN: | 1940-9818 |