Deformability Assessment of Waterborne Protozoa Using a Microfluidic-Enabled Force Microscopy Probe

Many modern filtration technologies are incapable of the complete removal of Cryptosporidium oocysts from drinking-water. Consequently, Cryptosporidium-contaminated drinking-water supplies can severely implicate both water utilities and consumers. Existing methods for the detection of Cryptosporidium in drinking-water do not discern between non-pathogenic and pathogenic species, nor between viable and non-viable oocysts. Using FluidFM, a novel force spectroscopy method employing microchannelled cantilevers for single-cell level manipulation, we assessed the size and deformability properties of two species of Cryptosporidium that pose varying levels of risk to human health. A comparison of such characteristics demonstrated the ability of FluidFM to discern between Cryptosporidium muris and Cryptosporidium parvum with 86% efficiency, whilst using a measurement throughput which exceeded 50 discrete oocysts per hour. In addition, we measured the deformability properties for untreated and temperature-inactivated oocysts of the highly infective, human pathogenic C. parvum to assess whether deformability may be a marker of viability. Our results indicate that untreated and temperature-inactivated C. parvum oocysts had overlapping but significantly different deformability distributions.

[1]  Hertz On the Contact of Elastic Solids , 1882 .

[2]  Kehe Huang,et al.  Comparison of viability and infectivity of Cryptosporidium parvum oocysts stored in potassium dichromate solution and chlorinated tap water. , 2007, Veterinary parasitology.

[3]  J. Rose,et al.  Drinking water treatment processes for removal of Cryptosporidium and Giardia. , 2004, Veterinary parasitology.

[4]  Tomaso Zambelli,et al.  Bacterial adhesion force quantification by fluidic force microscopy. , 2015, Nanoscale.

[5]  J. Sader,et al.  Calibration of rectangular atomic force microscope cantilevers , 1999 .

[6]  K. Hensel Journal für die reine und angewandte Mathematik , 1892 .

[7]  Huw Smith,et al.  Cryptosporidium excystation and invasion: getting to the guts of the matter. , 2005, Trends in parasitology.

[8]  J. Harris,et al.  Cryptosporidium parvum: structural components of the oocyst wall. , 1999, The Journal of parasitology.

[9]  C. Drummond,et al.  Oocysts of Cryptosporidium parvum and model sand surfaces in aqueous solutions: an atomic force microscope (AFM) study. , 2002, Water research.

[10]  B. Tl,et al.  Investigation of the interaction force between Cryptosporidium parvum oocysts and solid surfaces. , 2007 .

[11]  D. Dixon,et al.  Force of Interaction between a Biocolloid and an Inorganic Oxide: Complexity of Surface Deformation, Roughness, and Brushlike Behavior , 2001 .

[12]  J. Walz,et al.  Investigation of the interaction force between Cryptosporidium parvum oocysts and solid surfaces. , 2007, Langmuir.

[13]  Journal für die reine und angewandte Mathematik , 1893 .

[14]  H. Smith,et al.  Identification of Cryptosporidium Species and Genotypes in Scottish Raw and Drinking Waters during a One-Year Monitoring Period , 2010, Applied and Environmental Microbiology.

[15]  H. Dupont,et al.  Virulence of three distinct Cryptosporidium parvum isolates for healthy adults. , 1999, The Journal of infectious diseases.

[16]  Nicole K Henderson-Maclennan,et al.  Deformability-based cell classification and enrichment using inertial microfluidics. , 2011, Lab on a chip.

[17]  Tomaso Zambelli,et al.  Force-controlled manipulation of single cells: from AFM to FluidFM. , 2014, Trends in biotechnology.

[18]  J. Bates,et al.  Evaluation of Five Membrane Filtration Methods for Recovery of Cryptosporidium and Giardia Isolates from Water Samples , 2004, Applied and Environmental Microbiology.

[19]  Hansen Bow,et al.  Microfluidics for cell separation , 2010, Medical & Biological Engineering & Computing.

[20]  B. Tl,et al.  Interaction Force Profiles between Cryptosporidium parvum Oocysts and Silica Surfaces , 2005 .

[21]  H. Butt,et al.  Tilt of atomic force microscope cantilevers: effect on spring constant and adhesion measurements. , 2004, Langmuir : the ACS journal of surfaces and colloids.

[22]  C. Speer,et al.  Ultrastructure of Cryptosporidium parvum oocysts and excysting sporozoites as revealed by high resolution scanning electron microscopy. , 1985, The Journal of protozoology.

[23]  H. Hertz Ueber die Berührung fester elastischer Körper. , 1882 .

[24]  S. Enomoto,et al.  Cryptosporidium and cryptosporidiosis. , 2005, Advances in parasitology.

[25]  Tomaso Zambelli,et al.  FluidFM: combining atomic force microscopy and nanofluidics in a universal liquid delivery system for single cell applications and beyond. , 2009, Nano letters.

[26]  Han Wei Hou,et al.  Deformability based cell margination--a simple microfluidic design for malaria-infected erythrocyte separation. , 2010, Lab on a chip.

[27]  Tomaso Zambelli,et al.  Force-controlled patch clamp of beating cardiac cells. , 2015, Nano letters.

[28]  H Bridle,et al.  Deterministic lateral displacement for particle separation: a review. , 2014, Lab on a chip.

[29]  C. Drummond,et al.  Multi-scale Cryptosporidium/sand interactions in water treatment. , 2006, Water research.

[30]  R. Chalmers,et al.  Looking for Cryptosporidium: the application of advances in detection and diagnosis , 2013, Trends in Parasitology.

[31]  P. G. de Gennes,et al.  Polymers at an interface; a simplified view , 1987 .