Optimising single cell activity assessment of Lactobacillus plantarum by fluorescent in situ hybridisation as affected by growth.
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.
Wageningen Centre for Food Sciences, P.O. Box 557, 6700 AN Wageningen, The Netherlands. Maaike.deVries@wur.nl
Maaike C
MC
de Vries
Elaine E
EE
Vaughan
Michiel
M
Kleerebezem
Willem M
WM
de Vos
D002463:Cell Membrane Permeability; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007778:Lactobacillus / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D015345:Oligonucleotide Probes / Q000737:chemistry; D012336:RNA, Ribosomal, 16S / Q000737:chemistry / Q000235:genetics
D015345:Oligonucleotide Probes; D012336:RNA, Ribosomal, 16S