Nucleic acid stains as indicators of Giardia muris viability following cyst inactivation

Taghi-Kilani, R.; Gyürék, Lyndon L.; Millard, Paul J.; Finch, Gordon R.; Belosevic, Miodrag

doi:10.1016/0020-7519(96)00033-1

Cited by 33 publications

(17 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, before a more widespread acceptance of cytometric profiling as a measure of cell viability occurs, studies such as this must be undertaken to determine the extent to which various FCM profiles reflect the ability of cells to grow on microbial agar plates. Studies have previously shown these limitations (22,27). Hence, FCM data must be related to those obtained by traditional microbiological methods, and therefore FACS studies are a key component in understanding the relationship between FCM and plate count enumeration techniques.…”

Section: Discussionmentioning

confidence: 99%

Responses of Escherichia coli, Listeria monocytogenes, and Staphylococcus aureus to Simulated Food Processing Treatments, Determined Using Fluorescence-Activated Cell Sorting and Plate Counting

Kennedy

Cronin

Wilkinson

2011

Appl Environ Microbiol

View full text Add to dashboard Cite

Three common food pathogenic microorganisms were exposed to treatments simulating those used in food processing. Treated cell suspensions were then analyzed for reduction in growth by plate counting. Flow cytometry (FCM) and fluorescence-activated cell sorting (FACS) were carried out on treated cells stained for membrane integrity (Syto 9/propidium iodide) or the presence of membrane potential [DiOC 2 (3)]. For each microbial species, representative cells from various subpopulations detected by FCM were sorted onto selective and nonselective agar and evaluated for growth and recovery rates. In general, treatments giving rise to the highest reductions in counts also had the greatest effects on cell membrane integrity and membrane potential. Overall, treatments that impacted cell membrane permeability did not necessarily have a comparable effect on membrane potential. In addition, some bacterial species with extensively damaged membranes, as detected by FCM, appeared to be able to replicate and grow after sorting. Growth of sorted cells from various subpopulations was not always reflected in plate counts, and in some cases the staining protocol may have rendered cells unculturable. Optimized FCM protocols generated a greater insight into the extent of the heterogeneous bacterial population responses to food control measures than did plate counts. This study underlined the requirement to use FACS to relate various cytometric profiles generated by various staining protocols with the ability of cells to grow on microbial agar plates. Such information is a prerequisite for more-widespread adoption of FCM as a routine microbiological analytical technique.

show abstract

Section: Discussionmentioning

confidence: 99%

Responses of Escherichia coli, Listeria monocytogenes, and Staphylococcus aureus to Simulated Food Processing Treatments, Determined Using Fluorescence-Activated Cell Sorting and Plate Counting

Kennedy

Cronin

Wilkinson

2011

Appl Environ Microbiol

View full text Add to dashboard Cite

show abstract

“…The pathogen state information can include simple measures of viability to sophisticated measures of functional or metabolic abilities of the detected pathogen, all of which have been performed using fluorescent molecular probes (21)(22)(23)(24). It can also be used for more complicated measures of potential virulence that can be used to assess overall health dangers to humans or animals.…”

Section: Design Constraints For a Field Ready Pathogen Detectormentioning

confidence: 99%

A microfluidic-based hybrid SPR/molecular imaging biosensor for the multiplexed detection of foodborne pathogens

et al. 2009

View full text Add to dashboard Cite

It is important to screen our food supply for pathogen contaminations. Current methods to screen for bacterial contamination involve using costly reagents such as antibodies or PCR reagents or time-costly growth in cultures. There is need for portable, real-time, multiplex pathogen detection technology that can predict the safety of food where it is produced or distributed.Surface plasmon resonance (SPR) imaging is a sensitive, label-free method that can detect the binding of an analyte to a surface due to changes in refractive index that occur upon binding. It can be used for label-free detection of the presence of potential pathogens. Simultaneous fluorescence molecular imaging on the other side of the biochip can be used to ascertain pathogen status or functional state which may affect its potential danger to humans or animals.We are designing and testing hybrid microfluidic biochips to detect multiple pathogens using a combination of SPRI and fluorescence imaging. The device consists of an array of gold spots, each functionalized with a peptide targeting a specific pathogen. This peptide biosensor array is enclosed by a PDMS microfluidic flow chamber that delivers a magnetically concentrated sample to be tested. An SPR image is taken from the bottom of the biochip. Image analysis is used to quantify the amount of pathogen (both live and dead) bound to each spot. Since PDMS is very transmissive to visible light, an epi-fluorescence image is taken from the top of the biochip. Fluorescence imaging determines the live:dead ratio of each pathogen using an inexpensive SYTO 9®-Propidium Iodide assay. The volume of sample that the biochip can analyze is small, so possible pathogens are pre-concentrated using immunomagnetic separation. Functionalized magnetic particles are bound to pathogens present in the sample, and a magnet is used to separate them from the bulk fluid.

show abstract

“…SYTO ® 9 was developed by Molecular Probes and was offered in the live/dead BactLight kit together with propidium iodide (PI) and stains both live and dead cells. To our knowledge the first publication on SYTO ® 9 was by Taghi‐Kilani et al from 1996 in which SYTO ® 9 and the SYTO green colors ranging from number 11–16 were tested in relation to parasite viability. The first article in relation to bacteria is from 1999 by Boulos et al .…”

Section: Bacteria Biofilm and Chronic Bacterial Infectionsmentioning

confidence: 99%

The use of fluorescent staining techniques for microscopic investigation of polymorphonuclear leukocytes and bacteria

Alhede

Stavnsbjerg

Bjarnsholt

2018

APMIS

View full text Add to dashboard Cite

The use of fluorescent stains to visually investigate eukaryotic and/or prokaryotic cells is increasing quickly and manuscripts within all areas of research publish results using fluorescent staining techniques. However, in contrast to literature on traditional histological staining techniques, the literature on fluorescent stains and staining techniques does not offer as good an illustration of cellular morphology. The aim of this guideline is to illustrate different fluorescent stains and staining techniques for imaging immune cells, in particular the polymorphonuclear leukocytes (PMNs), in combination with infecting bacteria as seen in chronic bacterial infections. Thereby providing the first guideline for the morphology and identification of fluorescently stained PMNs, bacteria and biofilm.

show abstract

Nucleic acid stains as indicators of Giardia muris viability following cyst inactivation

Cited by 33 publications

References 26 publications

Responses of Escherichia coli, Listeria monocytogenes, and Staphylococcus aureus to Simulated Food Processing Treatments, Determined Using Fluorescence-Activated Cell Sorting and Plate Counting

Responses of Escherichia coli, Listeria monocytogenes, and Staphylococcus aureus to Simulated Food Processing Treatments, Determined Using Fluorescence-Activated Cell Sorting and Plate Counting

A microfluidic-based hybrid SPR/molecular imaging biosensor for the multiplexed detection of foodborne pathogens

The use of fluorescent staining techniques for microscopic investigation of polymorphonuclear leukocytes and bacteria

Contact Info

Product

Resources

About