Flow Cytometry Combined With Single Cell Sorting to Study Heterogeneous Germination of Bacillus Spores Under High Pressure

Zhang, Yifan; Delbrück, Alessia I.; Off, Cosima L.; Benke, Stephan; Mathys, Alexander

doi:10.3389/fmicb.2019.03118

Cited by 17 publications

(31 citation statements)

References 68 publications

(122 reference statements)

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“…a Understand the interaction of the probe and the micro-organism being studied and recognizing the growth stage of the micro-organism b Include sufficient negative and positive control to subtract any background fluorescence, including autofluorescence from the micro-organism itself (Zhang et al 2020). c Have a multiple staining and counterstaining strategy to triangulate results.…”

Section: Resultsmentioning

confidence: 99%

“…As highlighted by Magge et al (2009), autofluorescence may occur in some species as a consequence of spore coat components such as dityrosine which cross-links through oxidation activities involving peroxidase and oxidase. Hence, in the design of a FCM experiment, it may be necessary to consider the addition of a negative control for autofluorescence as adopted by Zhang et al (2020) in the execution of their research.…”

Section: Light Scattermentioning

confidence: 99%

“…Hence, in the design of a FCM experiment, it may be necessary to consider the addition of a negative control for autofluorescence as adopted by Zhang et al . (2020) in the execution of their research.…”

Section: Flow Cytometrymentioning

confidence: 99%

“…The very recent work of Zhang et al . (2020) demonstrates how FCM can be employed to elucidate various phenotypic fractions observed during an inactivation treatment. FCM can also show the sequence of events from intact spores to germinated spores (high fluorescence with Syto) to an intermediate (moderate fluorescence) ‘unknown state’ to finally an inactivated state (PI positive).…”

Section: Challenges With Fcm In Sterilization Microbiologymentioning

confidence: 99%

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Opportunities for the application of real‐time bacterial cell analysis using flow cytometry for the advancement of sterilization microbiology

McEvoy¹,

Lynch

Rowan

2020

J Appl Microbiol

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Medical devices provide critical care and diagnostic applications through patient contact. Sterility assurance level (SAL) may be defined as the probability of a single viable microorganism occurring on an item after a sterilization process. Sterilization microbiology often relies upon using an overkill validation method where a 12-log reduction in recalcitrant bacterial endospore population occurs during the process that exploits conventional laboratory-based culture media for enumeration. This timely review explores key assumptions underpinning use of conventional culture-based methods in sterilization microbiology. Consideration is given to how such methods may limit the ability to fully appreciate the inactivation kinetics of a sterilization process such as vaporized hydrogen peroxide (VH2O2) sterilization, and consequently design efficient sterilization processes. Specific use of the real-time flow cytometry (FCM) is described by way of elucidating the practical relevance of these limitation factors with implications and opportunities for the sterilization industry discussed. Application of FCM to address these culture-based limitation factors will inform real-time kinetic inactivation modelling and unlock potential to embrace emerging opportunities for pharma, medical device and sterilization industries including potentially disruptive applications that may involve reduced usage of sterilant.

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Section: Resultsmentioning

confidence: 99%

Section: Light Scattermentioning

confidence: 99%

Section: Flow Cytometrymentioning

confidence: 99%

Section: Challenges With Fcm In Sterilization Microbiologymentioning

confidence: 99%

See 2 more Smart Citations

Opportunities for the application of real‐time bacterial cell analysis using flow cytometry for the advancement of sterilization microbiology

McEvoy¹,

Lynch

Rowan

2020

J Appl Microbiol

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“…Furthermore, an unclear allocation could relate to changes in the metabolic state of cells that lead to a shift in their location on the density plots obtained from FCM. For instance, bacterial sporulation, such as that reported for species of the order Bacillales , increases the level of dye uptake, resulting in higher emission on the respective channel ( Zhang et al, 2020 ). Members affiliated with the order of Bacillales were also reported for microalgal cultures ( Steichen et al, 2020 ).…”

Section: Discussionmentioning

confidence: 99%

Automated Online Flow Cytometry Advances Microalgal Ecosystem Management as in situ, High-Temporal Resolution Monitoring Tool

Haberkorn

Off

Besmer³

et al. 2021

Front. Bioeng. Biotechnol.

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Microalgae are emerging as a next-generation biotechnological production system in the pharmaceutical, biofuel, and food domain. The economization of microalgal biorefineries remains a main target, where culture contamination and prokaryotic upsurge are main bottlenecks to impair culture stability, reproducibility, and consequently productivity. Automated online flow cytometry (FCM) is gaining momentum as bioprocess optimization tool, as it allows for spatial and temporal landscaping, real-time investigations of rapid microbial processes, and the assessment of intrinsic cell features. So far, automated online FCM has not been applied to microalgal ecosystems but poses a powerful technology for improving the feasibility of microalgal feedstock production through in situ, real-time, high-temporal resolution monitoring. The study lays the foundations for an application of automated online FCM implying far-reaching applications to impel and facilitate the implementation of innovations targeting at microalgal bioprocesses optimization. It shows that emissions collected on the FL1/FL3 fluorescent channels, harnessing nucleic acid staining and chlorophyll autofluorescence, enable a simultaneous assessment (quantitative and diversity-related) of prokaryotes and industrially relevant phototrophic Chlorella vulgaris in mixed ecosystems of different complexity over a broad concentration range (2.2–1,002.4 cells ⋅μL–1). Automated online FCM combined with data analysis relying on phenotypic fingerprinting poses a powerful tool for quantitative and diversity-related population dynamics monitoring. Quantitative data assessment showed that prokaryotic growth phases in engineered and natural ecosystems were characterized by different growth speeds and distinct peaks. Diversity-related population monitoring based on phenotypic fingerprinting indicated that prokaryotic upsurge in mixed cultures was governed by the dominance of single prokaryotic species. Automated online FCM is a powerful tool for microalgal bioprocess optimization owing to its adaptability to myriad phenotypic assays and its compatibility with various cultivation systems. This allows advancing bioprocesses associated with both microalgal biomass and compound production. Hence, automated online FCM poses a viable tool with applications across multiple domains within the biobased sector relying on single cell–based value chains.

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Fc-specific and covalent conjugation of a fluorescent protein to a native antibody through a photoconjugation strategy for fabrication of a novel photostable fluorescent antibody

Gao

et al. 2020

Anal Bioanal Chem

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Flow Cytometry Combined With Single Cell Sorting to Study Heterogeneous Germination of Bacillus Spores Under High Pressure

Cited by 17 publications

References 68 publications

Opportunities for the application of real‐time bacterial cell analysis using flow cytometry for the advancement of sterilization microbiology

Opportunities for the application of real‐time bacterial cell analysis using flow cytometry for the advancement of sterilization microbiology

Automated Online Flow Cytometry Advances Microalgal Ecosystem Management as in situ, High-Temporal Resolution Monitoring Tool

Fc-specific and covalent conjugation of a fluorescent protein to a native antibody through a photoconjugation strategy for fabrication of a novel photostable fluorescent antibody

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