Ca2+ in Hybridization Solutions for Fluorescence in situ Hybridization Facilitates the Detection of Enterobacteriaceae

Microbes and Environments
Shin HarutaYasuo Igarashi

Abstract

Fluorescence in situ hybridization (FISH) has been employed to identify microorganisms at the single cell level under a microscope. Extensive efforts have been made to improve and extend the FISH technique; however, the development of a widely applicable protocol is a continuing challenge. The present study evaluated the effects of divalent cations in the hybridization solution on the FISH-based detection of various species of bacteria and archaea with rRNA-targeted probes. A flow cytometric analysis after FISH with a standard hybridization buffer detected positive signals from less than 30% of Escherichia coli IAM 1264 cells. However, the number of cells with positive signals increased to more than 90% after the addition of calcium chloride to the hybridization buffer. Mn2+ also had positive effects, whereas Mg2+ did not. The positive effects of Ca2+ were similarly observed for bacteria belonging to Enterobacteriaceae, including Enterobacter sakazakii IAM 12660T, E. aerogenes IAM 12348, Klebsiella planticola IAM 14202, and Salmonella enterica subsp. enterica serovar Typhimurium strain LT2. These results indicate that the supplementation of Ca2+ to the hybridization buffer for FISH contributes to the efficient detection of Ente...Continue Reading

References

Dec 1, 1991·Journal of General Microbiology·B ZardaK H Schleifer
Jun 5, 1983·Journal of Molecular Biology·D Hanahan
Sep 1, 1994·Applied and Environmental Microbiology·S BurggrafK O Stetter
Jan 1, 1993·Annual Review of Microbiology·R KolterA Tormo
Jul 3, 1999·Nucleic Acids Research·S NakanoN Sugimoto
Jun 18, 2002·Journal of Applied Microbiology·M OotsuboH Oyaizu
Dec 3, 2004·Applied and Environmental Microbiology·L Safak Yilmaz, Daniel R Noguera
Jan 5, 2006·Applied and Environmental Microbiology·L Safak YilmazDaniel R Noguera
Oct 1, 1996·Applied and Environmental Microbiology·L RaskinD A Stahl
Feb 28, 2007·Biophysical Journal·Zhi-Jie Tan, Shi-Jie Chen
Apr 17, 2008·Nature Reviews. Microbiology·Rudolf Amann, Bernhard M Fuchs
Feb 13, 2010·Cold Spring Harbor Protocols·Annelie Wendeberg
Dec 17, 2011·Journal of Microbiological Methods·Shuji KawakamiKengo Kubota
Apr 8, 2014·Systematic and Applied Microbiology·Kengo KubotaFumio Inagaki
Jun 14, 2014·Frontiers in Microbiology·Gabriele BergKornelia Smalla
Jul 15, 2015·The ISME Journal·Martin KaltenpothAleš Svatoš
Jul 29, 2015·Systematic and Applied Microbiology·Tsuyoshi YamaguchiTakashi Yamaguchi
Feb 3, 2016·Nature Reviews. Microbiology·Naomi Attar

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Methods Mentioned

BETA
imaging techniques
flow cytometry

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