Fluorescent in situ hybridization en suspension (FISHES) using digoxigenin-labeled probes and flow cytometry.
Fluorescent in situ hybridization has become a major technique for visualizing genetic material in fixed cells. Currently, many systems utilize the hybridization of labeled molecular probes to cells that are attached to slides. We have developed a technique that allows for in situ hybridization to be performed using cells in suspension. By using digoxigenin-labeled DNA probes and a fluoresceinated antibody directed against the digoxigenin, we can measure the resulting signal on a flow cytometer and the cells can be attached to microscope slides for visual analysis.
Timm
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Department of Flow Cytometry, Roswell Park Cancer Institute, Buffalo, NY 14263.
Stewart
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D000818:Animals; D001483:Base Sequence; D001709:Biotechnology; D004247:DNA / Q000096:biosynthesis / Q000235:genetics; D004076:Digoxigenin; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D015335:Molecular Probes; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification
D015335:Molecular Probes; D012333:RNA, Messenger; D004247:DNA; D004076:Digoxigenin