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Updated: Jul 16, 2026
Studying Cryptosporidium Infection in 3D Tissue-derived Human Organoid Culture Systems by Microinjection
Published on: September 14, 2019
Quantitative tracking of Cryptosporidium infection in cell culture with CFSE
Hanping Feng1, Weijia Nie, Ruben Bonilla
1Division of Infectious Diseases, Tufts University, Cummings School of Veterinary Medicine, North Grafton, Massachusetts 01536, USA.
Abstract:
Immunofluorescence-based assays have been developed to detect and quantitate Cryptosporidium parvum infection in cell culture. Here, we describe a method that tracks and quantifies the early phase of attachment and invasion of C. parvum sporozoites using a fluorescent dye. Newly excysted sporozoites were labeled with the amine-reactive fluorescein probe carboxyfluorescein diacetate succinimidyl esters (CFSE) using an optimized protocol. The initial invasion of cells by labeled parasites was detected with fluorescent or confocal microscopy. The infection of cells was quantified by flow cytometry. Comparative analysis of infection of cells with CFSE-labeled and unlabeled sporozoites showed that the infectivity of C. parvum was not affected by CFSE labeling. Quantitative analysis showed that C. parvum Iowa and MD isolates were considerably more invasive than Cryptosporidium hominis isolate TU502. Unlike immunofluorescent assays, CFSE labeling permitted the tracking of the initial invasion of C. parvum. Such an assay may be useful for studying the dynamics of host cell-parasite interaction and possibly for drug screening.
Insights
Researchers developed a new method using fluorescent dye (CFSE) to track Cryptosporidium parvum invasion in cells. This technique accurately quantifies parasite attachment and invasion dynamics, aiding in host-parasite interaction studies.
Area of Science:
- Parasitology
- Cell Biology
- Microscopy and Imaging
Background:
- Immunofluorescence assays are used for Cryptosporidium parvum detection.
- A need exists to track early parasite attachment and invasion dynamics.
Purpose of the Study:
- To develop and validate a method for tracking and quantifying early-stage Cryptosporidium parvum sporozoite attachment and invasion.
- To compare the invasiveness of different Cryptosporidium isolates.
Main Methods:
- Newly excysted Cryptosporidium parvum sporozoites were labeled with carboxyfluorescein diacetate succinimidyl esters (CFSE).
- Parasite invasion was visualized using fluorescent and confocal microscopy.
- Infection levels were quantified via flow cytometry.
Main Results:
- CFSE labeling did not affect the infectivity of Cryptosporidium parvum sporozoites.
- CFSE labeling enabled tracking of initial parasite invasion, unlike traditional immunofluorescence assays.
- Cryptosporidium parvum isolates (Iowa, MD) demonstrated higher invasiveness compared to Cryptosporidium hominis (TU502).
Conclusions:
- CFSE labeling provides a robust method for studying early host cell-parasite interactions in Cryptosporidium infections.
- This assay is valuable for understanding invasion dynamics and potential drug screening for cryptosporidiosis.
