Home › Microbiology › Obtaining Highly Purified Toxoplasma gondii Oocysts by a Discontinuous Cesium Chloride Gradient
Microbiology JoVE (Open Access) Citable · DOI

Obtaining Highly Purified Toxoplasma gondii Oocysts by a Discontinuous Cesium Chloride Gradient

DOI: 10.3791/1420-v
What you'll learn
  • ✓ Purify T. gondii oocysts from cat feces using discontinuous CsCl gradient centrifugation
  • ✓ Apply sucrose flotation and density separation techniques for parasite isolation
  • ✓ Implement biosafety protocols when handling infectious T. gondii oocysts
  • ✓ Obtain purified oocysts suitable for molecular and tissue culture studies
Protocol

This study describes the development of a modified CsCl method that easily purifies T. gondii oocysts from feces of infected cats that are suitable for molecular biological and tissue culture manipulation

Difficulty
advanced
Total time
~4-6 hours per sample (fecal extraction through purification)
Biosafety
BSL-2

Steps

1
Review safety protocols for T. gondii oocyst handling

Understand and implement biosafety measures specific to T. gondii oocysts, including appropriate personal protective equipment and containment procedures before beginning work.

▶ 01:35
2
Prepare fecal sample and perform sucrose flotation

Extract and homogenize infected cat feces, then perform sucrose flotation to separate oocysts from fecal debris as an initial purification step.

▶ 03:15
3
Build and run discontinuous CsCl density gradient

Construct a cesium chloride density gradient and centrifuge the flotation product to further separate and purify oocysts based on buoyant density.

▶ 07:27
4
Collect and characterize purified oocyst fraction

Recover the purified oocyst band from the gradient and assess purity and morphology for suitability in downstream molecular or tissue culture applications.

▶ 10:03
💬 Comments coming soon