Optimising single cell activity assessment of Lactobacillus plantarum by fluorescent in situ hybridisation as affected by growth

M.C. de Vries, E.E. Vaughan, M. Kleerebezem, W.M. de Vos

Research output: Contribution to journalArticleAcademicpeer-review

17 Citations (Scopus)

Abstract

Fluorescent in situ hybridisation (FISH) with a 16S ribosomal RNA (rRNA)-targeted oligonucleotide probe, Eub338, could be used to estimate the in situ activity of Lactobacillus plantarum WCFS1 in exponentially growing cells. However, L. plantarum is capable of growth to very high cell densities, and the properties of the L. plantarum cell envelope prevented effective entry of the fluorescent oligonucleotide probe into the cells at later stages of growth at high cell densities. Total rRNA measurements of cells isolated at different growth stages showed maximal amounts of RNA (8.77 +/- 0.8 fg) per cell at the early stationary phase and confirmed the effectiveness of FISH for accurate activity measurement in exponentially growing cells. (C) 2004 Elsevier B.V. All rights reserved.
Original languageEnglish
Pages (from-to)109-115
JournalJournal of Microbiological Methods
Volume59
Issue number1
DOIs
Publication statusPublished - 2004

Fingerprint

Lactobacillus plantarum
Fluorescence In Situ Hybridization
Growth
Oligonucleotide Probes
Cell Count
Ribosomal RNA
Fluorescent Dyes

Keywords

  • ribosomal-rna
  • salmonella-typhimurium
  • messenger-rna
  • protein

Cite this

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title = "Optimising single cell activity assessment of Lactobacillus plantarum by fluorescent in situ hybridisation as affected by growth",
abstract = "Fluorescent in situ hybridisation (FISH) with a 16S ribosomal RNA (rRNA)-targeted oligonucleotide probe, Eub338, could be used to estimate the in situ activity of Lactobacillus plantarum WCFS1 in exponentially growing cells. However, L. plantarum is capable of growth to very high cell densities, and the properties of the L. plantarum cell envelope prevented effective entry of the fluorescent oligonucleotide probe into the cells at later stages of growth at high cell densities. Total rRNA measurements of cells isolated at different growth stages showed maximal amounts of RNA (8.77 +/- 0.8 fg) per cell at the early stationary phase and confirmed the effectiveness of FISH for accurate activity measurement in exponentially growing cells. (C) 2004 Elsevier B.V. All rights reserved.",
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Optimising single cell activity assessment of Lactobacillus plantarum by fluorescent in situ hybridisation as affected by growth. / de Vries, M.C.; Vaughan, E.E.; Kleerebezem, M.; de Vos, W.M.

In: Journal of Microbiological Methods, Vol. 59, No. 1, 2004, p. 109-115.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Optimising single cell activity assessment of Lactobacillus plantarum by fluorescent in situ hybridisation as affected by growth

AU - de Vries, M.C.

AU - Vaughan, E.E.

AU - Kleerebezem, M.

AU - de Vos, W.M.

PY - 2004

Y1 - 2004

N2 - Fluorescent in situ hybridisation (FISH) with a 16S ribosomal RNA (rRNA)-targeted oligonucleotide probe, Eub338, could be used to estimate the in situ activity of Lactobacillus plantarum WCFS1 in exponentially growing cells. However, L. plantarum is capable of growth to very high cell densities, and the properties of the L. plantarum cell envelope prevented effective entry of the fluorescent oligonucleotide probe into the cells at later stages of growth at high cell densities. Total rRNA measurements of cells isolated at different growth stages showed maximal amounts of RNA (8.77 +/- 0.8 fg) per cell at the early stationary phase and confirmed the effectiveness of FISH for accurate activity measurement in exponentially growing cells. (C) 2004 Elsevier B.V. All rights reserved.

AB - Fluorescent in situ hybridisation (FISH) with a 16S ribosomal RNA (rRNA)-targeted oligonucleotide probe, Eub338, could be used to estimate the in situ activity of Lactobacillus plantarum WCFS1 in exponentially growing cells. However, L. plantarum is capable of growth to very high cell densities, and the properties of the L. plantarum cell envelope prevented effective entry of the fluorescent oligonucleotide probe into the cells at later stages of growth at high cell densities. Total rRNA measurements of cells isolated at different growth stages showed maximal amounts of RNA (8.77 +/- 0.8 fg) per cell at the early stationary phase and confirmed the effectiveness of FISH for accurate activity measurement in exponentially growing cells. (C) 2004 Elsevier B.V. All rights reserved.

KW - ribosomal-rna

KW - salmonella-typhimurium

KW - messenger-rna

KW - protein

U2 - 10.1016/j.mimet.2004.06.010

DO - 10.1016/j.mimet.2004.06.010

M3 - Article

VL - 59

SP - 109

EP - 115

JO - Journal of Microbiological Methods

JF - Journal of Microbiological Methods

SN - 0167-7012

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