Universal membrane-labeling combined with expression of Katushka far-red fluorescent protein enables non-invasive dynamic and longitudinal quantitative 3D dual-color fluorescent imaging of multiple bacterial strains in mouse intestine

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Penate-Medina , O , Tower , R J , Penate-Medina , T , Will , O , Saris , P E J , Suojanen , J , Sorsa , T , Huuskonen , L , Hiippala , K , Satokari , R , Gluer , C C , de Vos , W M & Reunanen , J 2019 , ' Universal membrane-labeling combined with expression of Katushka far-red fluorescent protein enables non-invasive dynamic and longitudinal quantitative 3D dual-color fluorescent imaging of multiple bacterial strains in mouse intestine ' , BMC Microbiology , vol. 19 , 167 . https://doi.org/10.1186/s12866-019-1538-z

Title: Universal membrane-labeling combined with expression of Katushka far-red fluorescent protein enables non-invasive dynamic and longitudinal quantitative 3D dual-color fluorescent imaging of multiple bacterial strains in mouse intestine
Author: Penate-Medina, Oula; Tower, Robert J.; Penate-Medina, Tuula; Will, Olga; Saris, Per E. J.; Suojanen, Juho; Sorsa, Timo; Huuskonen, Laura; Hiippala, Kaisa; Satokari, Reetta; Gluer, Claus C.; de Vos, Willem M.; Reunanen, Justus
Contributor: University of Helsinki, Department of Microbiology
University of Helsinki, HUS Head and Neck Center
University of Helsinki, Department of Oral and Maxillofacial Diseases
University of Helsinki, Immunobiology Research Program
University of Helsinki, Immunobiology Research Program
University of Helsinki, Reetta Maria Satokari / Principal Investigator
University of Helsinki, Director and Common Matters
University of Helsinki, Biocenter Oulu
Date: 2019-07-18
Number of pages: 9
Belongs to series: BMC Microbiology
ISSN: 1471-2180
URI: http://hdl.handle.net/10138/305362
Abstract: Background: The human gastrointestinal (GI) tract microbiota has been a subject of intense research throughout the 3rd Millennium. Now that a general picture about microbiota composition in health and disease is emerging, questions about factors determining development of microbiotas with specific community structures will be addressed. To this end, usage of murine models for colonization studies remains crucial. Optical in vivo imaging of either bioluminescent or fluorescent bacteria is the basis for non-invasive detection of intestinal colonization of bacteria. Although recent advances in in vivo fluorescence imaging have overcome many limitations encountered in bioluminescent imaging of intestinal bacteria, such as requirement for live cells, high signal attenuation and 2D imaging, the method is still restricted to bacteria for which molecular cloning tools are available. Results: Here, we present usage of a lipophilic fluorescent dye together with Katushka far-red fluorescent protein to establish a dual-color in vivo imaging system to monitor GI transit of different bacterial strains, suitable also for strains resistant to genetic labeling. Using this system, we were able to distinguish two different E. coli strains simultaneously and show their unique transit patterns. Combined with fluorescence molecular tomography, these distinct strains could be spatially and temporally resolved and quantified in 3D. Conclusions: Developed novel method for labeling microbes and identify their passage both temporally and spatially in vivo makes now possible to monitor all culturable bacterial strains, also those that are resistant to conventional genetic labeling.
Subject: Bacterial colonization
Fluorescence imaging
Katushka
Dual-color imaging
Three-dimensional
Fluorescence molecular tomography
Murine model
Far-red fluorescent protein
In vivo
Intestine
ESCHERICHIA-COLI
GUT MICROBIOTA
LACTOBACILLUS-PLANTARUM
LACTOCOCCUS-LACTIS
IN-VITRO
PERSISTENCE
DISEASE
DIET
1183 Plant biology, microbiology, virology
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