Anti-fading agents for confocal immunofluorescence: colocalization of nuclear polypeptides.
We evaluated the performance of four anti-fading agents during acquisition of multiple optical sections near the widest diameter of Drosophila accessory gland nuclei using indirect immunofluorescence and confocal laser scanning microscopy. Two commercially available agents, Vectashield and SlowFade showed anti-fading properties that alleviated fluorochrome fading associated with the acquisition of multiple fluorescent optical Z-series from a single specimen by a confocal laser scanning system. Using these reagents, we were able to colocalize polypeptides through immunostained whole Drosophila nuclei.
Berrios
M
M
University Microscopy Imaging Center, State University of New York at Stony Brook 11794-8088, USA.
Colflesh
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D000818:Animals; D002467:Cell Nucleus / Q000737:chemistry / Q000648:ultrastructure; D004264:DNA Topoisomerases, Type I / Q000032:analysis / Q000276:immunology; D004331:Drosophila melanogaster / Q000737:chemistry; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes / Q000737:chemistry*; D006657:Histones / Q000032:analysis / Q000276:immunology; D007700:Kinetics; D018613:Microscopy, Confocal / Q000379:methods*; D009687:Nuclear Proteins / Q000032:analysis*; D010455:Peptides / Q000032:analysis* / Q000276:immunology
D005456:Fluorescent Dyes; D006657:Histones; D009687:Nuclear Proteins; D010455:Peptides; D004264:DNA Topoisomerases, Type I; D016650:Fluorescein-5-isothiocyanate