Use of flow cytometric methods for single-cell analysis in environmental microbiology

Czechowska, Kamila; Johnson, David R.; Meer, Jan Roelof van der

doi:10.1016/j.mib.2008.04.006

Cited by 140 publications

(104 citation statements)

References 59 publications

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“…Both labeling/sorting approaches 337 and phenotypic description of isolates provide an opportunity to better understand 338 how microbial membership contributes to community properties ( Figure 2, Letter E or 339 F). Labeling and cell sorting approaches (e.g., fluorescent in situ hybridization (FISH) 340 coupled with flow cytometry cell sorting 38 , or immunocapture such as with 341 bromodeosyuridine, BrdU) 39 provide powerful tools to constrain the complexity of the 342 microbiome and directly test hypotheses that link membership to community 343 properties or microbial processes. Labeling and sorting techniques allow the cells 344 that can be targeted with a stain or other label to be separated from the broader 345 community and then assayed for membership or phenotypes such as activity or 346 biomass composition.…”

Section: Applying and Testing The Proposed Framework 334mentioning

confidence: 99%

Understanding How Microbiomes Influence The Systems They Inhabit: Moving From A Correlative To A Causal Research Framework

Hall

Bernhardt

Bier

et al. 2016

Preprint

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Translating the ever-increasing wealth of information on microbiomes (environment, host, or built environment) to advance the understanding of system-level processes is proving to be an exceptional research challenge. One reason for this challenge is that relationships between characteristics of microbiomes and the system-level processes they influence are often evaluated in the absence of a robust conceptual framework and reported without elucidating the underlying causal mechanisms. The reliance on correlative approaches limits the potential to expand the inference of a single relationship to additional systems and advance the field. We propose that research focused on how microbiomes influence the systems they inhabit should work within a common framework and target known microbial processes that contribute to the system-level processes of interest. Here we identify three distinct categories of microbiome characteristics (microbial processes, microbial community properties, and microbial membership) and propose a framework to empirically link each of these categories to each other and the broader system level processes they affect. We posit that it is particularly important to distinguish microbial community properties that can be predicted from constituent taxa (community aggregated traits) from and those properties that are currently unable to be predicted from constituent taxa (emergent properties). Existing methods in microbial ecology can be applied to more explicitly elucidate properties within each of these categories and connect these three categories of microbial characteristics with each other. We view this proposed framework, gleaned from a breadth of research on environmental microbiomes and ecosystem processes, as a promising pathway with the potential to advance discovery and understanding across a broad range of microbiome science.

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Section: Applying and Testing The Proposed Framework 334mentioning

confidence: 99%

Understanding How Microbiomes Influence The Systems They Inhabit: Moving From A Correlative To A Causal Research Framework

Hall

Bernhardt

Bier

et al. 2016

Preprint

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“…Combining the flow cytometry with different fluorescent dyes has become an useful approach for the study of activated sludge used in the wastewater treatment processes and for the microbiological analysis of drinking water (30)(31)(32)(33)(34). It is expected that flow cytometry will revolution the microbiology in the future through understanding the complexity of microbial ecosystems and how they work and will aid the researchers to isolate some microorganisms uncharacterized until now (34). The aim of this paper was to investigate the influence of the electromagnetic field on morpho-physiological parameters of some bacterial strains isolated from wastewater, assessed through flow cytometry.…”

Section: Introductionmentioning

confidence: 99%

The Influence of Electromagnetic Field on Morphophysiological Parameters of Bacterial Cells Evaluated Through Flow Cytometry

Radu¹,

Marinescu²,

Savin³

et al. 2016

Simi 2016

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Flow cytometry represents an important tool in environmental biology, and especially in cellular biology, through its ability to distinguish among different physiological states (viable, latent and non-viable). Flow cytometry combined with fluorescent markers, such as propidium iodide and ethidium bromide allows to perform rapid measurements on individual cells and also simultaneous measurements of multiple cellular parameters, both structural and functional. The aim of this paper was to investigate the influence of the electromagnetic field on the morpho-physiological parameters of some bacterial strains cells isolated from wastewater, assessed through flow cytometry. In this respect, a number of 10 bacterial strains isolated from wastewater (E. coli, Salmonella sp., Enterobacter sp., Citrobacter sp., Klebsiella sp. and Enterococcus sp.) were exposed to an electromagnetic field (50Hz electric field at different voltages) for 24 hours, at 37°C. Both samples, the ones exposed to electromagnetic field and blank samples (unexposed) were assessed by flow cytometry technique. Two possible mechanism of action have been tracked, i.e. the efflux pumps activity (fluorescence markerEthidium Bromide) and permeabilisation of cellular layers (fluorescence marker -Propidium Iodide). Both mechanisms of action have been identified, from slightly to significant modifications, as compared to untreated controls, quantified as ΔMFI (median of fluorescence intensity). Conclusion: Our results suggest that flow cytometry could be used for the real-time evaluation of the influence of different electromagnetic fields on the aquatic microbiota, the obtained results being of great interest for the development of technological solution with increased efficiency in the (waste)water treatment.

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“…Immunophenotyping is used extensively within hematology, and allows identification of multiple phenotypic and functional markers enabling the comprehensive interrogation of cells on a single cell level (Czechowska et al 2008;Shapiro 1883). The advantage of flow cytometry is the combination of multiplexed assays; measuring several features of a single cell simultaneously, whilst sampling a statistically significant number of cells (~5000).…”

Section: Introductionmentioning

confidence: 99%

Multiparameter flow cytometry for the characterization of human embryonic stem cells

et al. 2012

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Use of flow cytometric methods for single-cell analysis in environmental microbiology

Cited by 140 publications

References 59 publications

Understanding How Microbiomes Influence The Systems They Inhabit: Moving From A Correlative To A Causal Research Framework

Understanding How Microbiomes Influence The Systems They Inhabit: Moving From A Correlative To A Causal Research Framework

The Influence of Electromagnetic Field on Morphophysiological Parameters of Bacterial Cells Evaluated Through Flow Cytometry

Multiparameter flow cytometry for the characterization of human embryonic stem cells

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