The DOI matches its Crossref record.
Optimizing fluorescent in situ hybridization with rRNA‐targeted oligonucleotide probes for flow cytometric identification of microorganisms
Open accessAbstract
A combination of fluorescent rRNA‐targeted oligonucleotide probes (“phylogenetic stains”) and flow cytometry was used for a high resolution automated analysis of mixed microbial populations. Fixed cells of bacteria and yeasts were hybridized in suspension with fluorescein‐ or tetramethylrhodamine‐labeled oligonucleotide probes complementary to group‐specific regions of the 16S ribosomal RNA (rRNA) molecules. Quantifying probe‐conferred cell fluorescence by flow cytometry, we could discriminate between target and nontarget cell populations. We critically examined changes of the hybridization conditions, kinetics of the hybridization, and posthybridization treatments. Intermediate probe concentrations, addition of detergent to the hybridization buffer, and a posthybridization washing step were found to increase the signal to noise ratio. We could demonstrate a linear correlation between growth rate and probe‐conferred fluorescence of Escherichia coli and Pseudomonas cepacia cells. Oligonucleotides labeled with multiple fluorochromes showed elevated levels of non‐specific binding and therefore could not be used to lower the detection limits, which still restrict studies with fluorescing rRNA‐targeted oligonucleotide probes to well‐growing microbial cells. Two probes of different specificities—one labeled with fluorescein, the other with tetramethylrhodamine—could be applied simultaneously for dual color analysis. © 1993 Wiley‐Liss, Inc.
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Mueller, S., Saunier, K., Hanisch, C., Norin, E., Alm, L., Midtvedt, T., Cresci, A., Silvi, S., Orpianesi, C., Verdenelli, M. C., et al. (2006) · Applied and Environmental Microbiology · doi:10.1128/aem.72.2.1027-1033.2006 · Cited by 1,006
Absence of intestinal microbiota does not protect mice from diet-induced obesity
Fleissner, C. K., Huebel, N., Abd El-Bary, M. M., Loh, G., Klaus, S., & Blaut, M. (2010) · British Journal of Nutrition · doi:10.1017/s0007114510001303 · Cited by 436
Effects of a gluten-free diet on gut microbiota and immune function in healthy adult human subjects
De Palma, G., Nadal, I., Collado, M. C., & Sanz, Y. (2009) · British Journal Of Nutrition · doi:10.1017/s0007114509371767 · Cited by 344
De Palma, G., Nadal, I., Medina, M., Donat, E., Ribes-Koninckx, C., Calabuig, M., & Sanz, Y. (2010) · BMC Microbiology · doi:10.1186/1471-2180-10-63 · Cited by 330
Applications of flow cytometry to clinical microbiology
Álvarez-Barrientos, A., Arroyo, J., Cantón, R., Nombela, C., & Sánchez-Pérez, M. (2000) · Clinical Microbiology Reviews · doi:10.1128/cmr.13.2.167 · Cited by 300
Adipose tissue derived bacteria are associated with inflammation in obesity and type 2 diabetes
Massier, L., Chakaroun, R., Tabei, S., Crane, A., Didt, K. D., Fallmann, J., von Bergen, M., Haange, S.-B., Heyne, H., Stumvoll, M., et al. (2020) · Gut · doi:10.1136/gutjnl-2019-320118 · Cited by 194
Biedermann, L., Brülisauer, K., Zeitz, J., Frei, P., Scharl, M., Vavricka, S. R., Fried, M., Loessner, M. J., Rogler, G., & Schuppler, M. (2014) · Inflammatory Bowel Diseases · doi:10.1097/mib.0000000000000129 · Cited by 178
Grimaldi, R., Cela, D., Swann, J. R., Vulevic, J., Gibson, G. R., Tzortzis, G., & Costabile, A. (2017) · FEMS Microbiology Ecology · doi:10.1093/femsec/fiw233 · Cited by 109
Host Responses to Intestinal Microbial Antigens in Gluten-Sensitive Mice
Natividad, J. M., Huang, X., Slack, E., Jury, J., Sanz, Y., David, C., Denou, E., Yang, P., Murray, J., McCoy, K. D., et al. (2009) · PLoS ONE · doi:10.1371/journal.pone.0006472 · Cited by 73
Fleming, E. J., Davis, R. E., McAllister, S. M., Chan, C. S., Moyer, C. L., Tebo, B. M., & Emerson, D. (2013) · FEMS Microbiology Ecology · doi:10.1111/1574-6941.12104 · Cited by 72
Microbial diversity of garden snail mucus
Belouhova, M., Daskalova, E., Yotinov, I., Topalova, Y., Velkova, L., Dolashki, A., & Dolashka, P. (2022) · MicrobiologyOpen · doi:10.1002/mbo3.1263 · Cited by 21
Chlamydiae as symbionts of photosynthetic dinoflagellates
Maire, J., Collingro, A., Tandon, K., Jameson, V. J., Judd, L. M., Horn, M., Blackall, L. L., & van Oppen, M. J. H. (2024) · The ISME Journal · doi:10.1093/ismejo/wrae139 · Cited by 7
Metadata
- Authors
- Wallner, G., Amann, R., & Beisker, W.
- Venue
- Cytometry
- Publisher
- Wiley
- Published
- 1993-01
- Volume
- 14
- Issue
- 2
- Pages
- 136-143
- ISSN
- 0196-4763 (ISSN-L)
- 1097-0320
- OpenAlex
- W2035775871
- Cited by
- 1,212
- References
- 0
- FWCI
- 7.33
- Type
- journal article
- Language
- en
- Open access
- bronze
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