title: A simple protocol for attenuating the auto-fluorescence of cyanobacteria for optimized fluorescence in situ hybridization (FISH) imaging.
abstract: Cyanobacteria contain pigments, which generate auto-fluorescence that interferes with fluorescence in situ hybridization (FISH) imaging of cyanobacteria. We describe simple chemical treatments using CuSO4 or H2O2 that significantly reduce the auto-fluorescence of Microcystis strains. These protocols were successfully applied in FISH experiments using 16S rRNA specific probes and filamentous cyanobacteria.
authors.affiliation: INSERM U1193, Physiopathogénèse et Traitement des Maladies du Foie, Hôpital Paul Brousse, bâtiment Lavoisier, 14 avenue Paul Vaillant Couturier, 94807 Villejuif Cedex, France.
authors.forename: Perrine
authors.initials: P
authors.lastname: Zeller
authors.affiliation: Laboratoire des Energies de Demain, LIED, UMR 8236 CNRS, Université Paris Diderot-Paris 7, 75013 Paris, France; Chimie ParisTech, 75005 Paris, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Ploux
authors.affiliation: Laboratoire des Energies de Demain, LIED, UMR 8236 CNRS, Université Paris Diderot-Paris 7, 75013 Paris, France. Electronic address: annick.mejean@univ-paris-diderot.fr.
authors.forename: Annick
authors.initials: A
authors.lastname: Méjean
mesh_terms: D001431:Bacteriological Techniques / Q000379:methods*; D001483:Base Sequence; D019327:Copper Sulfate / Q000737:chemistry; D000458:Cyanobacteria / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D005453:Fluorescence; D006861:Hydrogen Peroxide / Q000494:pharmacology; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D046931:Microcystis / Q000187:drug effects / Q000235:genetics; D008856:Microscopy, Fluorescence; D015345:Oligonucleotide Probes / Q000235:genetics; D061848:Optical Imaging / Q000379:methods*; D010539:Permeability; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity
chemical_list: D015345:Oligonucleotide Probes; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D006861:Hydrogen Peroxide; D019327:Copper Sulfate
keywords: Auto-fluorescence; CuSO(4); Cyanobacteria; H(2)O(2); Microcystis; fluorescence in situ hybridization