Quantification of bacterial populations in complex ecosystems using fluorescent in situ hybridization, confocal laser scanning microscopy and image analysis

Autor: Théodore Bouchez, René Moletta, Michael Wagner, Patrick Dabert, Jean-Jacques Godon
Přispěvatelé: Laboratoire de Biotechnologie de l'Environnement [Narbonne] (LBE), Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro), Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut National de la Recherche Agronomique (INRA)
Jazyk: angličtina
Rok vydání: 2001
Předmět:
IV — Inventaire
Caractérisation et Suivi de la Diversité Microbienne / Inventory
Characterization and Monitoring of Microbial Diversity
Background fluorescence
In situ hybridization
analyse d'image
Biology
biofilm
law.invention
03 medical and health sciences
boue activée
hybridation fluorescente in situ
nitrospira
Confocal microscopy
law
Microscopy
Genetics
Confocal laser scanning microscopy
medicine
microbiologie
Ecology
Evolution
Behavior and Systematics
ComputingMilieux_MISCELLANEOUS
030304 developmental biology
bactérie
floc
0303 health sciences
BIOTECHNOLOGIE
[SDV.GEN]Life Sciences [q-bio]/Genetics
medicine.diagnostic_test
030306 microbiology
population microbienne
quantification bactérienne
caractérisation
General Medicine
Fluorescence
microscopie laser confocale
Surface measurement
nitrosomonas
Animal Science and Zoology
fluorescence
Biological system
nitrobacter
Fluorescence in situ hybridization
Zdroj: Genetics Selection Evolution
Genetics Selection Evolution, BioMed Central, 2001, 33 (Suppl1), pp.S307-S318
Scopus-Elsevier
Theodore Bouchez
Genetics Selection Evolution Suppl1 (33), S307-S318. (2001)
ResearcherID
ISSN: 0999-193X
1297-9686
Popis: A procedure for quantification of distinct bacterial populations in aggre-gated ecosystems was developed. It is based on fluorescent in situ hybridization coupled with confocal-laser-scanning microscopy and surface measurement of hybridized bacteria by image analysis. The proportion of a targeted bacterial species was ob-tained by comparison between the surface hybridized by a specific probe and the surface hybridized with a general bacterial reference probe. The accuracy of the re-sults obtained was evaluated by direct visual counting on the same sets of images. The analytical uncertainty of the image analysis procedure was determined and de-pended on the threshold values selected by the operator for cutting between signal and background fluorescence. The number of fields to be analyzed for reliable quantification was also determined. This procedure allows a quantification of the proportions of bacterial species in aggregated bacterial ecosystems which gives more accurate re-sults than visual counting because of the large number of bacteria counted. Moreover, since floc structures are preserved, it also gives information about the distribution of bacterial populations in the floc material which might reveal bacterial interactions within the community.
Databáze: OpenAIRE
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