Abstract:
:In this study, the feasibility of fluorescence lifetime imaging (FLIM) for measurement of RNA:DNA ratios in microorganisms was assessed. The fluorescence lifetime of a nucleic acid-specific probe (SYTO 13) was used to directly measure the RNA:DNA ratio inside living bacterial cells. In vitro, SYTO 13 showed shorter fluorescence lifetimes in DNA solutions than in RNA solutions. Growth experiments with bacterial monocultures were performed in liquid media. The results demonstrated the suitability of SYTO 13 for measuring the growth-phase-dependent RNA:DNA ratio in Escherichia coli cells. The fluorescence lifetime of SYTO 13 reflected the known changes of the RNA:DNA ratio in microbial cells during different growth phases. As a result, the growth rate of E. coli cells strongly correlated with the fluorescence lifetime. Finally, the fluorescence lifetimes of SYTO 13 in slow- and fast-growing biofilms were compared. For this purpose, biofilms developed from activated sludge were grown as autotrophic and heterotrophic communities. The FLIM data clearly showed a longer fluorescence lifetime for the fast-growing heterotrophic biofilms and a shorter fluorescence lifetime for the slow-growing autotrophic biofilms. Furthermore, starved biofilms showed shorter lifetimes than biofilms supplied with glucose, indicating a lower RNA:DNA ratio in starved biofilms. It is suggested that FLIM in combination with SYTO 13 represents a useful tool for the in situ differentiation of active and inactive bacteria. The technique does not require radioactive chemicals and may be applied to a broad range of sample types, including suspended and immobilized microorganisms.
journal_name
Appl Environ Microbioljournal_title
Applied and environmental microbiologyauthors
Walczysko P,Kuhlicke U,Knappe S,Cordes C,Neu TRdoi
10.1128/AEM.01806-07subject
Has Abstractpub_date
2008-01-01 00:00:00pages
294-9issue
1eissn
0099-2240issn
1098-5336pii
AEM.01806-07journal_volume
74pub_type
杂志文章abstract::Factors affecting the production of [14C]patulin from [1-14C]acetate by replacement cultures of Penicillium patulum have been investigated. Incorporation of [1-14C]acetate into patulin reached a maximum with 6- to 8-day-old cultures incubated at 28 degrees C for 8 h in a replacement medium containing 0.1 M glucose, in...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.33.4.814-816.1977
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.61.3.1150-1152.1995
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.59.11.3744-3749.1993
更新日期:1993-11-01 00:00:00
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pub_type: 杂志文章
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更新日期:2005-08-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.34.5.557-563.1977
更新日期:1977-11-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.54.2.588-589.1988
更新日期:1988-02-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.38.5.831-835.1979
更新日期:1979-11-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.61.11.3910-3918.1995
更新日期:1995-11-01 00:00:00
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journal_title:Applied and environmental microbiology
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doi:10.1128/AEM.53.5.1046-1050.1987
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pub_type: 杂志文章
doi:10.1128/AEM.60.6.2137-2146.1994
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.54.3.688-692.1988
更新日期:1988-03-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
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更新日期:2014-01-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.64.8.2914-2919.1998
更新日期:1998-08-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/aem.68.9.4534-4538.2002
更新日期:2002-09-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.03110-14
更新日期:2015-03-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.55.3.617-622.1989
更新日期:1989-03-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
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更新日期:2014-06-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.53.6.1277-1285.1987
更新日期:1987-06-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.01355-12
更新日期:2012-09-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.02987-13
更新日期:2014-01-01 00:00:00