Journal
JOURNAL OF BIOLOGICAL ENGINEERING
Volume 12, Issue -, Pages -Publisher
BIOMED CENTRAL LTD
DOI: 10.1186/s13036-018-0095-6
Keywords
Cryptosporidium parvum oocysts; Disposable microfluidic micromixers; Imaging flow cytometry; Water quality; Immunocytochemistry; Fluorescence microscopy
Funding
- NHMRC [APP1045841]
- NORTE-45-2015-02 program [NORTE-01-0145-FEDER-000029]
- NHMRC CDA Fellowship
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Background: Protecting drinking water supplies from pathogens such as Cryptosporidium parvum is a major concern for water utilities worldwide. The sensitivity and specificity of current detection methods are largely determined by the effectiveness of the concentration and separation methods used. The purpose of this study is to develop micromixers able to specifically isolate and concentrate Cryptosporidium, while allowing in situ analysis. Results: In this study, disposable microfluidic micromixers were fabricated to effectively isolate Cryptosporidium parvum oocysts from water samples, while allowing direct observation and enabling quantification of oocysts captured in the device using high quality immunofluorescence microscopy. In parallel, quantitative analysis of the capture yield was carried out by analyzing the waste from the microfluidics outlet with an Imaging Flow Cytometer. At the optimal flow rate, capture efficiencies up to 96% were achieved in spiked samples. Conclusions: Scaled microfluidic isolation and detection of Cryptosporidium parvum will provide a faster and more efficient detection method for Cryptosporidium compared to other available laboratory-scale technologies.
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