The Application of Multi-Parameter Flow Cytometry to Monitor Individual Microbial Cell Physiological State

2004 ◽  
pp. 197-223 ◽  
Author(s):  
Christopher J. Hewitt ◽  
Gerhard Nebe-Von-Caron
Keyword(s):  
Flow Cytometry ◽  
Physiological State ◽  
Microbial Cell

Biocalorimetry and laser flow cytometry for characterization of the physiological state of microorganisms

1996 ◽  
Vol 277 ◽  
pp. 7-16 ◽  
Author(s):  
Brigitte Zentgraf
Keyword(s):  
Flow Cytometry ◽  
Physiological State

scholarly journals Intracellular pH Distribution in Saccharomyces cerevisiae Cell Populations, Analyzed by Flow Cytometry

2005 ◽  
Vol 71 (3) ◽  
pp. 1515-1521 ◽  
Author(s):  
Minoska Valli ◽  
Michael Sauer ◽  
Paola Branduardi ◽  
Nicole Borth ◽  
Danilo Porro ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Flow Cytometry ◽  
Intracellular Ph ◽  
Environmental Changes ◽  
Physiological State ◽  
Yeast Cells ◽  
Cell Populations ◽  
Ph Homeostasis ◽  
Ph Distribution ◽  
External Ph

ABSTRACT Intracellular pH has an important role in the maintenance of the normal functions of yeast cells. The ability of the cell to maintain this pH homeostasis also in response to environmental changes has gained more and more interest in both basic and applied research. In this study we describe a protocol which allows the rapid determination of the intracellular pH of Saccharomyces cerevisiae cells. The method is based on flow cytometry and employs the pH-dependent fluorescent probe carboxy SNARF-4F. The protocol attempts to minimize the perturbation of the system under study, thus leading to accurate information about the physiological state of the single cell. Moreover, statistical analysis performed on major factors that may influence the final determination supported the validity of the optimized protocol. The protocol was used to investigate the effect of external pH on S. cerevisiae cells incubated in buffer. The results obtained showed that stationary cells are better able than exponentially grown cells to maintain their intracellular pH homeostasis independently of external pH changes. Furthermore, analysis of the intracellular pH distribution within the cell populations highlighted the presence of subpopulations characterized by different intracellular pH values. Notably, a different behavior was observed for exponentially grown and stationary cells in terms of the appearance and development of these subpopulations as a response to a changing external pH.

Assessment of physiological state of microorganisms in activated sludge with flow cytometry: application for monitoring sludge production minimization

2008 ◽  
Vol 35 (11) ◽  
pp. 1261-1268 ◽  
Author(s):  
A. Prorot ◽  
C. Eskicioglu ◽  
R. Droste ◽  
C. Dagot ◽  
P. Leprat
Keyword(s):  
Flow Cytometry ◽  
Activated Sludge ◽  
Physiological State ◽  
Sludge Production

Microbial cell budgets of an Arctic glacier surface quantified using flow cytometry

2012 ◽  
Vol 14 (11) ◽  
pp. 2998-3012 ◽  
Author(s):  
T. D. L. Irvine-Fynn ◽  
A. Edwards ◽  
S. Newton ◽  
H. Langford ◽  
S. M. Rassner ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Microbial Cell ◽  
Glacier Surface ◽  
Arctic Glacier

scholarly journals Monitoring the Physiological State in the Dark Fermentation of Maize/Grass Silage Using Flow Cytometry and Electrooptic Polarizability Measurements

2020 ◽  
Author(s):  
Carlos E. Gómez-Camacho ◽  
Klaus Pellicer Alborch ◽  
Anika Bockisch ◽  
Peter Neubauer ◽  
Stefan Junne ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Physiological State ◽  
Dark Fermentation ◽  
Grass Silage

scholarly journals Flow Cytometry to Assess the Counts and Physiological State of Cronobacter sakazakii Cells after Heat Exposure

Foods ◽  
2019 ◽  
Vol 8 (12) ◽  
pp. 688 ◽  
Author(s):  
Paloma Cal-Sabater ◽  
Irma Caro ◽  
María J. Castro ◽  
María J. Cao ◽  
Javier Mateo ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Physiological State ◽  
Heat Treatments ◽  
Cell Suspensions ◽  
Plate Count ◽  
Dead Cell ◽  
Cronobacter Sakazakii ◽  
Infant Formulae ◽  
Heat Treated ◽  
The Impact

Cronobacter sakazakii is an opportunistic pathogen that is associated with outbreaks of neonatal necrotizing enterocolitis, septicaemia, and meningitis. Reconstituted powdered infant formulae is the most common vehicle of infection. The aim of the present study is to gain insight into the physiological states of C. sakazakii cells using flow cytometry to detect the compromised cells, which are viable but non-culturable using plate-based methods, and to evaluate the impact of milk heat treatments on those populations. Dead-cell suspensions as well as heat-treated and non-heat-treated cell suspensions were used. After 60 or 65 °C treatments, the number of compromised cells increased as a result of cells with compromised membranes shifting from the heat-treated suspension. These temperatures were not effective at killing all bacteria but were effective at compromising their membranes. Thus, mild heat treatments are not enough to guarantee the safety of powered infant formulae. Flow cytometry was capable of detecting C. sakazakii’s compromised cells that cannot be detected with classical plate count methods; thus, it could be used as a screening test to decrease the risk derived from the presence of pathogenic viable but non-culturable cells in this food that is intended for newborns’ nutrition.

scholarly journals High Throughput Volume Flow Cytometry (parallel-iLIFE) Resolves Mitochondrial Network On the Go

2020 ◽  
Author(s):  
Prashant Kumar ◽  
Prakash Joshi ◽  
Jigmi Basumatary ◽  
Partha Pratim Mondal
Keyword(s):  
Flow Cytometry ◽  
High Throughput ◽  
Microfluidic Chip ◽  
Physiological State ◽  
Volume Visualization ◽  
Light Sheet ◽  
Disease Diagnosis ◽  
Volume Flow ◽  
Critical Parameters ◽  
Mitochondrial Network

Cell screening and viability studies are paramount to access cell morphology and intracellular molecular variations within large heterogeneous populations of cells. This forms the basis for diagnosis of infections, evaluating immunohistochemistry and routine histopathology. The proposed volume flow cytometry (also termed as, parallel Integrated Light-sheet imaging and flow-based enquiry (parallel-iLIFE)) is a powerful method that adds new capabilities (3D volume visualization, organelle-level resolution and multi-organelle screening) powered by light sheet based illumination. Unlike state-of-the-art point-illumination based imaging cytometry techniques, light sheet based parallel-iLIFE technique is capable of screening species with high throughput and near diffraction-limited resolution. The flow system was realized on a multichannel (Y-type) microfluidic chip that enables visualization of mitochondrial network of several cells in-parallel at a relatively high flow-rate of 2000 nl/min. The calibration of system requires study of point emitters (fluorescent beads) at physiologically relevant flow-rates (50−2000 nl/min) for determining flow-induced optical aberration in the system point spread function (PSF). Subsequently, recorded raw images and volumes were deconvolved with flow-variant PSF to reconstruct cellular mitochondrial network. High throughput investigation of HeLa cells were carried out at sub-cellular resolution in real-time and critical parameters (mitochondria count and size distribution, morphology and cell strain statistics) are determined on-the-go. These parameters determine the physiological state of cells and the changes in mitochondrial distribution over-time that may have consequences in disease diagnosis. The development of volume flow cytometry system (parallel-iLIFE) and its suitability to study sub-cellular components at high-throughput high-content capacity with organelle-level resolution may enable disease diagnosis on a single microfluidic chip.

scholarly journals Bacterial Flow Cytometry and Imaging as Potential Process Monitoring Tools for Industrial Biotechnology

Fermentation ◽  
2020 ◽  
Vol 6 (1) ◽  
pp. 10 ◽  
Author(s):  
Sumana Kadamalakunte Narayana ◽  
Sanjaya Mallick ◽  
Henrik Siegumfeldt ◽  
Frans van den Berg
Keyword(s):  
Flow Cytometry ◽  
Physiological State ◽  
Cluster Formation ◽  
Membrane Integrity ◽  
Process Variations ◽  
Industrial Biotechnology ◽  
Cell Size Distribution ◽  
Industrial Processing ◽  
Imaging Results ◽  
Visual Confirmation

Minimizing process variations by early identification of deviations is one approach to make industrial production processes robust. Cell morphology is a direct representation of the physiological state and an important factor for the cell’s survival in harsh environments as encountered during industrial processing. The adverse effects of fluctuating process parameters on cells were studied using flow cytometry and imaging. Results showed that altered pH caused a shift in cell size distribution from a heterogeneous mix of elongated and short cells to a homogenous population of short cells. Staining based on membrane integrity revealed a dynamics in the pattern of cluster formation during fermentation. Contradictory findings from forward scatter and imaging highlight the need for use of complementary techniques that provide visual confirmation to interpret changes. An atline flow cytometry or imaging capable of identifying subtle population deviations serves as a powerful monitoring tool for industrial biotechnology.

SYBR green as a fluorescent probe to evaluate the biofilm physiological state of Staphylococcus epidermidis, using flow cytometry

2011 ◽  
Vol 57 (10) ◽  
pp. 850-856 ◽  
Author(s):  
Filipe Cerca ◽  
Gabriela Trigo ◽  
Alexandra Correia ◽  
Nuno Cerca ◽  
Joana Azeredo ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Staphylococcus Epidermidis ◽  
Physiological State ◽  
Staining Intensity ◽  
Sybr Green ◽  
Tetrazolium Chloride ◽  
Viable But Nonculturable ◽  
Bacterial Rna ◽  
Nonculturable Cells ◽  
Excess Glucose

Staphylococcus epidermidis biofilms with different proportions of viable but nonculturable bacteria were used to show that SYBR green (SYBR) may be used as a probe to evaluate the bacterial physiological state using flow cytometry. Biofilms grown in excess glucose presented significantly higher proportions of dormant bacteria than biofilms grown in excess glucose with buffered pH conditions or with exponential-phase planktonic cultures. Bacteria obtained from biofilms with high or low proportions of viable but nonculturable cells were further cultured in broth medium and stained with SYBR at different time points. An association between bacterial growth and SYBR staining intensity was observed. In addition, bacteria presenting higher SYBR fluorescence intensity also stained more intensely with cyanoditolyl tetrazolium chloride, used as a probe to evaluate cellular metabolism. Accordingly, planktonic bacteria treated with rifampicin, an inhibitor of bacterial RNA transcription, presented lower SYBR and cyanoditolyl tetrazolium chloride staining intensity than nontreated bacteria. Overall, our results indicate that SYBR, in addition to being used as a component of LIVE/DEAD stain, may also be used as a probe to evaluate the physiological state of S. epidermidis cells.

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