Methodology and application of flow cytometry for investigation of human malaria parasites

Abstract

Historically, examinations of the inhibition of malaria parasite growth/invasion, whether using drugs or antibodies, have relied on the use of microscopy or radioactive hypoxanthine uptake. These are considered gold standards for measuring the effectiveness of antimalarial treatments, however, these methods have well known shortcomings. With the advent of flow cytometry coupled with the use of fluorescent DNA stains allowed for increased speed, reproducibility, and qualitative estimates of the effectiveness of antibodies and drugs to limit malaria parasite growth which addresses the challenges of traditional techniques. Because materials and machines available to research facilities are so varied, different methods have been developed to investigate malaria parasites by flow cytometry. This review is intended to serve as a reference guide for advanced users and importantly, as a primer for new users, to support expanded use and improvements to malaria flow cytometry, particularly in endemic countries.

Protective Effect of Chronic Schistosomiasis in Baboons Coinfected with Schistosoma mansoni and Plasmodium knowlesi

“…To record parasitemia, RBCs were stained using ethidium bromide as previously described (38). DiIC1-5 (1,1=,3,3,3=,3=-hexamethylindodicarbocyanine iodide) was included in the staining protocol for the detection of live parasites (39). Parasitemia was read at 0 and 48 h on a Becton-Dickinson LSRII flow cytometer (Franklin Lakes, NJ).…”

Advances in Antimalarial Drug Evaluation and New Targets for Antimalarials

“…Flow cytometry is currently used to study the biology of malaria parasites, but is still little used to evaluate parasite drug susceptibility, certainly because of the high cost of cytometers and a capacity restricted to moderate throughput assays (Grimberg, 2011). However, owing to the large range of fluorescent probes available having different biological or biochemical affinities that can be simultaneously analysed, flow cytometry allows the access to many more information than just parasite proliferation (DNA content).…”

Comparative studies of serum-free media and detection techniques for <i>in vitro</i> drug sensitivity assessment of <i>Plasmodium falciparum</i>

“…FCM has a particular advantage in determining populations of dividing stages of the parasite. FCM that uses different intercalating dyes has already been used successfully to test human and murine samples [14,20,21]. However, some of the dyes lack sufficient sensitivity and/or require complicated preparation procedures that make their use problematic.…”