Flowcytometric and ImageStream Rna-Fish Gene Expression, Quantification and Phenotypic Characterization of Blood Sporozoites and Sporozoites From Human Malaria Species

Thanks to the combination of Plasmodium-specific RNA detection and surface and nucleus staining, the new Flow-FISH method allowed us to characterize and quantify asexual and sexual blood-stage parasite from P vivax clinical samples as well as liver-stage parasites. We adapted the RNA FISH Stellaris method to specifically detect the expression of Plasmodium genes by flow cytometry and ImageStream (Flow-FISH). This new method accurately quantified the erythrocytic forms of (1) Plasmodium falciparum and Plasmodium vivax and (2) the sexual stages of P vivax from patient isolates. ImageStream analysis of liver stage sporozoites using a combination of surface circumsporozoite protein (CSP), deoxyribonucleic acid, and 18S RNA labeling proved that the new Flow-FISH is suitable for gene expression studies of transmission stages. This powerful multiparametric single-cell method offers a platform of choice for both applied and fundamental research on the biology of malaria parasites.

Automated summary

The new Flow-FISH method combines RNA detection and cell staining techniques to accurately quantify different stages of malaria parasites, suitable for gene expression studies and analysis of liver-stage sporozoites.