Quantum Dots as alternatives to organic fluorophores for Cryptosporidium detection using conventional flow cytometry and specific monoclonal antibodies: lessons learned.
BACKGROUND
Significant developments in biological applications are occurring through the incorporation of Quantum Dots (QDs) as biological labels. The demonstration of QDs unique optical properties may have important implications for the study of environmental samples, where microorganisms of interest need to be isolated away from the background debris.


METHODS
Flow cytometric analysis was used to determine the fluorescence intensity of oocysts after mAb staining by QDs or organic fluorophore conjugates. In addition, the level of non-specific binding to detrital particles within a control water concentrate was estimated using the optimal staining concentration determined for each mAb analyzed.


RESULTS
Under 488 nm excitation, oocysts stained with QD-conjugates exhibited significantly lower fluorescence intensity than organic conjugates. Moreover, the level of non-specific binding by QD-conjugates to detrital particles present in the water concentrate was significantly higher that of the organic conjugates.


CONCLUSIONS
While QDs are noted for their superior spectral characteristics, they have been shown here to be unsuitable for conventional flow cytometric detection of Cryptosporidium. Therefore, we conclude that in their current form, QD's are severely limited for fluorescent detection of pathogens in environmental applications.
Ferrari
B C
BC
Department of Chemistry and Biomolecular Sciences, Environmental Biotechnology CRC and Biotechnology Research Institute, Macquarie University Sydney, New South Wales 2109, Australia. bferrari@rna.bio.mq.edu.au
Bergquist
P L
PL
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D016785:Cryptosporidium parvum / Q000276:immunology / Q000302:isolation & purification*; D005434:Flow Cytometry / Q000379:methods*; D005455:Fluorescent Antibody Technique / Q000379:methods*; D005456:Fluorescent Dyes; D033661:Oocysts / Q000378:metabolism; D045663:Quantum Dots
D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes