Use of viability staining in combination with flow cytometry for rapid viability assessment of Lactobacillus rhamnosus GG in complex protein matrices

Journal of Microbiological Methods
S B DohertyA Brodkorb

Abstract

The aim of this study was to demonstrate that flow cytometry (FACS) could potentially be employed for rapid viability assessment of probiotic bacteria immobilized or encapsulated in complex matrices. Lactobacillus rhamnosus GG was immobilized within six different protein environments using whey protein isolate (WPI) and yoghurt matrices and encapsulated within protein micro-beads, all of which ranged in structural complexity. Following a series of environmental-stress trials, survival of the strain was examined using FACS compared to traditional plate count techniques. Cell extraction and digestive pre-treatments were designed to release cells and reduce the protein background, respectively, which represent compositional obstacles for efficient FACS analysis. Physico-chemical properties of protein-probiotic components revealed the mechanism necessary for efficient cell delivery during FACS analysis. This assay required 40 min sample preparation and distinct functional populations were discriminated based on fluorescent properties of thiazole orange (TO) and propidium iodide (PI). This assay yielded 45-50 samples/h, a detection range of 10(2)-10(10)cfu/ml of homogenate and generated correlation coefficients (r) of 0.95, 0.92 and...Continue Reading

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Citations

Jun 27, 2014·The British Journal of Nutrition·Louise M SullivanAndré Brodkorb
Aug 3, 2014·International Journal of Food Microbiology·Danyue Zhao, Nagendra P Shah
Jul 17, 2012·Journal of Microbiological Methods·Julie MonthéardSandrine Alfenore
Mar 6, 2013·European Journal of Pharmaceutical Sciences : Official Journal of the European Federation for Pharmaceutical Sciences·F Martin-DejardinO Chambin
Aug 18, 2017·Cytometry. Part B, Clinical Cytometry·Anne H TavenierPaul H P Groeneveld
May 16, 2018·Journal of Clinical Gastroenterology·Marco PaneLuca Mogna
Mar 18, 2015·Applied Microbiology and Biotechnology·Saúl AlonsoMario Díaz
Sep 16, 2020·Soft Matter·Faustine GomandClaire Gaiani

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