Using flow cytometry to detect protozoa

Hoffman, Rebecca M.; Standridge, Jon H.; Prieve, Audrey F.; Cucunato, Joseph C.; Bernhardt, Mat

doi:10.1002/j.1551-8833.1997.tb08293.x

Cited by 22 publications

(5 citation statements)

References 12 publications

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“…Cryptosporidium and Giardia vary in size and can be differentiated by their light‐scatter properties 13 , 20 . During analysis, a region was drawn around each population, and the fluorescence intensity of the organisms in each region was determined on a log 90 degree light scatter versus log green fluorescence histogram.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of four commercial antibodies

Hoffman¹,

Chauret²,

Standridge³

et al. 1999

Journal AWWA

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Evaluation of commercially available anti‐Cryptosporidium and anti‐Giardia antibodies indicates each offers advantages and disadvantages. Flow cytometer‐derived fluorescence measurements were used to evaluate lot‐to‐lot variability, cross‐reactivity, and avidity of four antibodies. Variability within kit 3 and 4 antibody lots was great, whereas kits 1 and 2 stained parasites consistently. Kit 3 and 4 antibodies cross‐reacted with non‐parvum Cryptosporidium; kit 1 and 4 antibodies cross‐reacted with Giardia muris. Kit 1 (primary only), 2, and 3 anti‐Cryptosporidium antibodies formed low‐avidity complexes, unlike those in kits 1 (secondary) and 4 and all Giardia antibodies. Additionally, the authors quantified the ability of antibodies to bind stressed and unstressed parasites following methodological processing. The studies identified significant differences in antibody characteristics and performance.

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Section: Methodsmentioning

confidence: 99%

Evaluation of four commercial antibodies

Hoffman¹,

Chauret²,

Standridge³

et al. 1999

Journal AWWA

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“…Flow cytometry has the ability to measure and analyze various physical properties of a single cell, especially the ova/cysts when they pass via a light beam [93]. The differentiation of cells is based on the relative size, granularity and the intensity of fluorescence [94].…”

Section: Flow Cytometrymentioning

confidence: 99%

A Review on the Current Knowledge and Prospects for the Development of Improved Detection Methods for Soil-Transmitted Helminth Ova for the Safe Reuse of Wastewater and Mitigation of Public Health Risks

Ravindran

Soni

Ball

2019

Water

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Climate change, increase in population and scarcity of freshwater have led to a global demand for wastewater reuse in irrigation. However, wastewater has to be treated in order to minimize the presence of pathogens, in particular, the ova of soil-transmitted helminthes (STHs). Limiting the transmission via removal of STH ova, accurate assessment of risks and minimizing the exposure to the public have been recommended by health regulators. The World Health Organization (WHO) guideline specifies a limit of ≤1 ova/L for safe wastewater reuse. Additionally, the Australian Guidelines for Water recycling (AGWR) recommend a hydraulic retention time of over 25 days in a lagoon or stabilization pond to ensure a 4 log reduction value of helminth ova and to mitigate soil-transmitted helminths associated risks to humans. However, the lack of fast and sensitive methods for assessing the concentration of STH ova in wastewater poses a considerable challenge for an accurate risk assessment. Consequently, it has been difficult to control soil-transmitted helminthiasis despite effective mass drug administration. This limitation can be overcome with the advent of novel techniques for the detection of helminth ova. Therefore, this review presents an assessment of the current methods to detect the viable ova of soil-transmitted helminths in wastewater. Furthermore, the review focuses on the perspectives for the emerging state-of-the-art research and developments that have the potential to replace currently available conventional and polymerase chain reaction based methods and achieve the guidelines of the WHO in order to allow the safe reuse of wastewater for non-potable applications, thereby minimizing public health risks.

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“…This method only requires basic equipment and a dozen microliters of cell culture but has a disadvantage of being low throughput, prone to human errors and subject to high variation. Meanwhile, advanced machineries, such as the Coulter counter, flow cytometer, image cytometer and microfluidic cytometer, have been applied to quantify cell numbers [ 8 , 9 ]. These expensive and specialized devices, however, may not be accessible for laboratories with limited resources or under an educational setting.…”

Section: Introductionmentioning

confidence: 99%

Performance Comparison of Five Methods for Tetrahymena Number Counting on the ImageJ Platform: Assessing the Built-in Tool and Machine-Learning-Based Extension

Kurnia

Sampurna²,

Audira³

et al. 2022

IJMS

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Previous methods to measure protozoan numbers mostly rely on manual counting, which suffers from high variation and poor efficiency. Although advanced counting devices are available, the specialized and usually expensive machinery precludes their prevalent utilization in the regular laboratory routine. In this study, we established the ImageJ-based workflow to quantify ciliate numbers in a high-throughput manner. We conducted Tetrahymena number measurement using five different methods: particle analyzer method (PAM), find maxima method (FMM), trainable WEKA segmentation method (TWS), watershed segmentation method (WSM) and StarDist method (SDM), and compared their results with the data obtained from the manual counting. Among the five methods tested, all of them could yield decent results, but the deep-learning-based SDM displayed the best performance for Tetrahymena cell counting. The optimized methods reported in this paper provide scientists with a convenient tool to perform cell counting for Tetrahymena ecotoxicity assessment.

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Using flow cytometry to detect protozoa

Cited by 22 publications

References 12 publications

Evaluation of four commercial antibodies

Evaluation of four commercial antibodies

A Review on the Current Knowledge and Prospects for the Development of Improved Detection Methods for Soil-Transmitted Helminth Ova for the Safe Reuse of Wastewater and Mitigation of Public Health Risks

Performance Comparison of Five Methods for Tetrahymena Number Counting on the ImageJ Platform: Assessing the Built-in Tool and Machine-Learning-Based Extension

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