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Appl Environ Microbiol. May 1996; 62(5): 1649–1655.
PMCID: PMC167939

Application of the novel nucleic acid dyes YOYO-1, YO-PRO-1, and PicoGreen for flow cytometric analysis of marine prokaryotes.

Abstract

Novel blue light-excited fluorescent dyes for nucleic acids (YOYO-1, YO-PRO-1, and PicoGreen) were tested on cultures of Escherichia coli and of a variety of marine prokaryotes. Results of flow cytometric DNA analyses were compared with those obtained with the UV-excited dyes bis-benzimide Hoechst 33342 or 4', 6-diamidino-2-phenylindole (DAPI). YOYO-1, YO-PRO-1, and PicoGreen can be used only on aldehyde-fixed cells and need to be supplemented with cofactors such as potassium, citrate, or EDTA. They are highly sensitive to ionic strength. Consequently, seawater culture samples cannot be stained directly with these dyes and require at least a 10-fold dilution with distilled water to obtain reliable fluorescence signals. After treatment with RNase, coefficients of variation for the G1 peak of the DNA distributions of the different strains tested with YOYO-1 or PicoGreen indicated in general an improvement over Hoechst 33342 staining. These novel dyes can be used to enumerate prokaryotic cells by flow cytometry, as demonstrated with E. coli. However, their sensitivity to ionic strength makes them unsuitable for cell cycle analysis in natural samples.

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Selected References

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  • Armbrust EV, Bowen JD, Olson RJ, Chisholm SW. Effect of light on the cell cycle of a marine synechococcus strain. Appl Environ Microbiol. 1989 Feb;55(2):425–432. [PMC free article] [PubMed]
  • Binder BJ, Chisholm SW. Relationship between DNA cycle and growth rate in Synechococcus sp. strain PCC 6301. J Bacteriol. 1990 May;172(5):2313–2319. [PMC free article] [PubMed]
  • Binder BJ, Chisholm SW. Cell Cycle Regulation in Marine Synechococcus sp. Strains. Appl Environ Microbiol. 1995 Feb;61(2):708–717. [PMC free article] [PubMed]
  • Button DK, Robertson BR. Kinetics of bacterial processes in natural aquatic systems based on biomass as determined by high-resolution flow cytometry. Cytometry. 1989 Sep;10(5):558–563. [PubMed]
  • Glazer AN, Rye HS. Stable dye-DNA intercalation complexes as reagents for high-sensitivity fluorescence detection. Nature. 1992 Oct 29;359(6398):859–861. [PubMed]
  • Hess WR, Weihe A, Loiseaux-de Goër S, Partensky F, Vaulot D. Characterization of the single psbA gene of Prochlorococcus marinus CCMP 1375 (Prochlorophyta). Plant Mol Biol. 1995 Mar;27(6):1189–1196. [PubMed]
  • Hirons GT, Fawcett JJ, Crissman HA. TOTO and YOYO: new very bright fluorochromes for DNA content analyses by flow cytometry. Cytometry. 1994 Feb 1;15(2):129–140. [PubMed]
  • Kepner RL, Jr, Pratt JR. Use of fluorochromes for direct enumeration of total bacteria in environmental samples: past and present. Microbiol Rev. 1994 Dec;58(4):603–615. [PMC free article] [PubMed]
  • Lebaron P, Joux F. Flow cytometric analysis of the cellular DNA content of Salmonella typhimurium and Alteromonas haloplanktis during starvation and recovery in seawater. Appl Environ Microbiol. 1994 Dec;60(12):4345–4350. [PMC free article] [PubMed]
  • Monger BC, Landry MR. Flow cytometric analysis of marine bacteria with hoechst 33342. Appl Environ Microbiol. 1993 Mar;59(3):905–911. [PMC free article] [PubMed]
  • Ong LJ, Glazer AN, Waterbury JB. An unusual phycoerythrin from a marine cyanobacterium. Science. 1984 Apr 6;224(4644):80–83. [PubMed]
  • Pinder AC, Purdy PW, Poulter SA, Clark DC. Validation of flow cytometry for rapid enumeration of bacterial concentrations in pure cultures. J Appl Bacteriol. 1990 Jul;69(1):92–100. [PubMed]
  • Potin P, Sanseau A, Le Gall Y, Rochas C, Kloareg B. Purification and characterization of a new kappa-carrageenase from a marine Cytophaga-like bacterium. Eur J Biochem. 1991 Oct 1;201(1):241–247. [PubMed]
  • Rye HS, Yue S, Wemmer DE, Quesada MA, Haugland RP, Mathies RA, Glazer AN. Stable fluorescent complexes of double-stranded DNA with bis-intercalating asymmetric cyanine dyes: properties and applications. Nucleic Acids Res. 1992 Jun 11;20(11):2803–2812. [PMC free article] [PubMed]
  • Skarstad K, Steen HB, Boye E. Cell cycle parameters of slowly growing Escherichia coli B/r studied by flow cytometry. J Bacteriol. 1983 May;154(2):656–662. [PMC free article] [PubMed]
  • Vaulot D, Courties C, Partensky F. A simple method to preserve oceanic phytoplankton for flow cytometric analyses. Cytometry. 1989 Sep;10(5):629–635. [PubMed]
  • Vaulot D, Marie D, Olson RJ, Chisholm SW. Growth of prochlorococcus, a photosynthetic prokaryote, in the equatorial pacific ocean. Science. 1995 Jun 9;268(5216):1480–1482. [PubMed]
  • Wyman M, Gregory RP, Carr NG. Novel Role for Phycoerythrin in a Marine Cyanobacterium, Synechococcus Strain DC2. Science. 1985 Nov 15;230(4727):818–820. [PubMed]

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