Imaging Flow Cytometry Demonstrates Physiological and Morphological Diversity within Treated Probiotic Bacteria Groups
Probiotic bacteria can be introduced to stresses during the culturing phase as an alternative to the use of protectants and coating substances during drying. Accurate enumeration of the bacterial count in a probiotic formulation can be provided using imaging flow cytometry (IFC). IFC overcomes the w...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-04-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/7/6841 |
| _version_ | 1797607667921846272 |
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| author | Jakub Kiepś Wojciech Juzwa Radosław Dembczyński |
| author_facet | Jakub Kiepś Wojciech Juzwa Radosław Dembczyński |
| author_sort | Jakub Kiepś |
| collection | DOAJ |
| description | Probiotic bacteria can be introduced to stresses during the culturing phase as an alternative to the use of protectants and coating substances during drying. Accurate enumeration of the bacterial count in a probiotic formulation can be provided using imaging flow cytometry (IFC). IFC overcomes the weak points of conventional, commonly used flow cytometry by combining its statistical power with the imaging content of microscopy in one system. Traditional flow cytometers only collect the fluorescence signal intensities, while IFC provides many more steps as it correlates the data on the measured parameters of fluorescence light with digitally processed images of the analyzed cells. As an alternative to standard methods (plate cell counts and traditional flow cytometry) IFC provides additional insight into the physiology and morphology of the cell. The use of complementary dyes (RedoxSensorTM Green and propidium iodide) allows for the designation of groups based on their metabolic activity and membrane damage. Additionally, cell sorting is incorporated to assess each group in terms of growth on different media (MRS-Agar and MRS broth). Results show that the groups with intermediate metabolic activity and some degree of cellular damage correspond with the description of viable but nonculturable cells. |
| first_indexed | 2024-03-11T05:34:04Z |
| format | Article |
| id | doaj.art-aa8755a98c8d481daaa2c43956cca8a5 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T05:34:04Z |
| publishDate | 2023-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-aa8755a98c8d481daaa2c43956cca8a52023-11-17T16:56:47ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-04-01247684110.3390/ijms24076841Imaging Flow Cytometry Demonstrates Physiological and Morphological Diversity within Treated Probiotic Bacteria GroupsJakub Kiepś0Wojciech Juzwa1Radosław Dembczyński2Department of Biotechnology and Food Microbiology, Poznan University of Life Sciences, Wojska Polskiego 48, 60-627 Poznań, PolandDepartment of Biotechnology and Food Microbiology, Poznan University of Life Sciences, Wojska Polskiego 48, 60-627 Poznań, PolandDepartment of Biotechnology and Food Microbiology, Poznan University of Life Sciences, Wojska Polskiego 48, 60-627 Poznań, PolandProbiotic bacteria can be introduced to stresses during the culturing phase as an alternative to the use of protectants and coating substances during drying. Accurate enumeration of the bacterial count in a probiotic formulation can be provided using imaging flow cytometry (IFC). IFC overcomes the weak points of conventional, commonly used flow cytometry by combining its statistical power with the imaging content of microscopy in one system. Traditional flow cytometers only collect the fluorescence signal intensities, while IFC provides many more steps as it correlates the data on the measured parameters of fluorescence light with digitally processed images of the analyzed cells. As an alternative to standard methods (plate cell counts and traditional flow cytometry) IFC provides additional insight into the physiology and morphology of the cell. The use of complementary dyes (RedoxSensorTM Green and propidium iodide) allows for the designation of groups based on their metabolic activity and membrane damage. Additionally, cell sorting is incorporated to assess each group in terms of growth on different media (MRS-Agar and MRS broth). Results show that the groups with intermediate metabolic activity and some degree of cellular damage correspond with the description of viable but nonculturable cells.https://www.mdpi.com/1422-0067/24/7/6841viabilityfluid bed dryinglactic acid bacteriastress factorsquality controlrapid assessment |
| spellingShingle | Jakub Kiepś Wojciech Juzwa Radosław Dembczyński Imaging Flow Cytometry Demonstrates Physiological and Morphological Diversity within Treated Probiotic Bacteria Groups International Journal of Molecular Sciences viability fluid bed drying lactic acid bacteria stress factors quality control rapid assessment |
| title | Imaging Flow Cytometry Demonstrates Physiological and Morphological Diversity within Treated Probiotic Bacteria Groups |
| title_full | Imaging Flow Cytometry Demonstrates Physiological and Morphological Diversity within Treated Probiotic Bacteria Groups |
| title_fullStr | Imaging Flow Cytometry Demonstrates Physiological and Morphological Diversity within Treated Probiotic Bacteria Groups |
| title_full_unstemmed | Imaging Flow Cytometry Demonstrates Physiological and Morphological Diversity within Treated Probiotic Bacteria Groups |
| title_short | Imaging Flow Cytometry Demonstrates Physiological and Morphological Diversity within Treated Probiotic Bacteria Groups |
| title_sort | imaging flow cytometry demonstrates physiological and morphological diversity within treated probiotic bacteria groups |
| topic | viability fluid bed drying lactic acid bacteria stress factors quality control rapid assessment |
| url | https://www.mdpi.com/1422-0067/24/7/6841 |
| work_keys_str_mv | AT jakubkieps imagingflowcytometrydemonstratesphysiologicalandmorphologicaldiversitywithintreatedprobioticbacteriagroups AT wojciechjuzwa imagingflowcytometrydemonstratesphysiologicalandmorphologicaldiversitywithintreatedprobioticbacteriagroups AT radosławdembczynski imagingflowcytometrydemonstratesphysiologicalandmorphologicaldiversitywithintreatedprobioticbacteriagroups |