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Dual staining of natural bacterioplankton with 4',6-diamidino-2-phenylindole and fluorescent oligonucleotide probes targeting kingdom-level 16S rRNA sequences
Open accessAbstract
A method for quantifying eubacterial cell densities in dilute communities of small bacterioplankton is presented. Cells in water samples were stained with 4',6-diamidino-2-phenylindole (DAPI), transferred to gelatin-coated slides, and hybridized with rhodamine-labeled oligonucleotide probes specific for kingdom-level 16S rRNA sequences. Between 48 and 69% of the cells captured on membrane filters were transferred to gelatin-coated slides. The number of DAPI-stained cells that were visualized with eubacterial probes varied from 35 to 67%. Only 2 to 4% of these cells also fluoresced following hybridization with a probe designed to target a eukaryotic 16S rRNA sequence. Between 0.1 and 6% of the bacterioplankton in these samples were autofluorescent and may have been mistaken as cells that hybridized with fluorescent oligonucleotide probes. Dual staining allows precise estimates of the efficiency of transfers of cells to gelatin films and can be used to measure the percentage of the total bacterioplankton that also hybridize with fluorescent oligonucleotide probes, indicating specific phylogenetic groups.
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Cited by
Suau, A., Bonnet, R., Sutren, M., Godon, J.-J., Gibson, G. R., Collins, M. D., & Doré, J. (1999) · Applied and Environmental Microbiology · doi:10.1128/aem.65.11.4799-4807.1999 · Cited by 1,490
SAR11 clade dominates ocean surface bacterioplankton communities
Morris, R. M., Rappé, M. S., Connon, S. A., Vergin, K. L., Siebold, W. A., Carlson, C. A., & Giovannoni, S. J. (2002) · Nature · doi:10.1038/nature01240 · Cited by 1,129
Kassinen, A., Krogius-Kurikka, L., Mäkivuokko, H., Rinttilä, T., Paulin, L., Corander, J., Malinen, E., Apajalahti, J., & Palva, A. (2007) · Gastroenterology · doi:10.1053/j.gastro.2007.04.005 · Cited by 1,002
Hubert, C., & Voordouw, G. (2007) · Applied and Environmental Microbiology · doi:10.1128/aem.02332-06 · Cited by 316
Mühling, M., Woolven-Allen, J., Murrell, J. C., & Joint, I. (2008) · The ISME Journal · doi:10.1038/ismej.2007.97 · Cited by 304
Metadata
- Authors
- Hicks, R. E., Amann, R. I., & Stahl, D. A.
- Venue
- Applied and Environmental Microbiology
- Publisher
- American Society for Microbiology
- Published
- 1992-07
- Volume
- 58
- Issue
- 7
- Pages
- 2158-2163
- ISSN
- 0099-2240 (ISSN-L)
- 1098-5336
- OpenAlex
- W1859696357
- Cited by
- 321
- References
- 0
- FWCI
- 12.12
- Type
- journal article
- Language
- en
- Open access
- green
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- Hicks, R. E., Amann, R. I., & Stahl, D. A. (1992). Dual staining of natural bacterioplankton with 4',6-diamidino-2-phenylindole and fluorescent oligonucleotide probes targeting kingdom-level 16S rRNA sequences. Applied and Environmental Microbiology. https://doi.org/10.1128/aem.58.7.2158-2163.1992
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