Visualisation and quantitative analysis of the rodent malaria liver stage by real time imaging.

PloS One
Ivo H J PloemenBlandine M D Franke-Fayard

Abstract

The quantitative analysis of Plasmodium development in the liver in laboratory animals in cultured cells is hampered by low parasite infection rates and the complicated methods required to monitor intracellular development. As a consequence, this important phase of the parasite's life cycle has been poorly studied compared to blood stages, for example in screening anti-malarial drugs. Here we report the use of a transgenic P. berghei parasite, PbGFP-Luc(con), expressing the bioluminescent reporter protein luciferase to visualize and quantify parasite development in liver cells both in culture and in live mice using real-time luminescence imaging. The reporter-parasite based quantification in cultured hepatocytes by real-time imaging or using a microplate reader correlates very well with established quantitative RT-PCR methods. For the first time the liver stage of Plasmodium is visualized in whole bodies of live mice and we were able to discriminate as few as 1-5 infected hepatocytes per liver in mice using 2D-imaging and to identify individual infected hepatocytes by 3D-imaging. The analysis of liver infections by whole body imaging shows a good correlation with quantitative RT-PCR analysis of extracted livers. The luminescenc...Continue Reading

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Methods Mentioned

BETA
transgenic
flow cytometry
Assay
dissection
PCR
FACS
transfection
fluorescence-microscopy

Software Mentioned

Living Image
SPSS
Living Image®
GraphPad Prism
Lumina

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