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

PLoS One. 2016 Mar 3;11(3):e0150438. doi: 10.1371/journal.pone.0150438. eCollection 2016.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cryptosporidium / isolation & purification*
  • Cryptosporidium parvum / isolation & purification*
  • Drinking Water / parasitology*
  • Elasticity
  • Humans
  • Microfluidics / instrumentation
  • Microfluidics / methods*
  • Microscopy, Atomic Force / instrumentation
  • Microscopy, Atomic Force / methods*
  • Oocysts / chemistry
  • Single-Cell Analysis
  • Water Purification / instrumentation
  • Water Purification / methods

Substances

  • Drinking Water

Grants and funding

This work was supported by 1) Royal Academy of Engineering / Engineering and Physical Sciences Research Council (EPSRC) Fellowship—Dr. Helen Bridle—Grant number: EP/G058881/1 URL—https://www.epsrc.ac.uk/; 2) NanoNextNL—SL -Grant Number: 06C.17 URL—http://www.nanonextnl.nl/; and Science and Technology Facilities Council—JM—Award Number: 1444568 URL: http://www.stfc.ac.uk/. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.