Cell trace far-red is a suitable erythrocyte dye for multi-color Plasmodium falciparum invasion phenotyping assays
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Date
2020-01-05
Journal Title
Journal ISSN
Volume Title
Publisher
Experimental Biology and Medicine
Abstract
Plasmodium falciparum erythrocyte invasion phenotyping assays are a very useful tool for
assessing parasite diversity and virulence, and for characterizing the formation of ligand–
receptor interactions. However, such assays need to be highly sensitive and reproducible,
and the selection of labeling dyes for differentiating donor and acceptor erythrocytes is a
critical factor. We investigated the suitability of cell trace far-red (CTFR) as a dye for P.
falciparum invasion phenotyping assays. Using the dyes carboxyfluorescein diacetate succinimidyl
ester (CFDA-SE) and dichloro dimethyl acridin one succinimidyl ester (DDAO-SE)
as comparators, we used a dye-dilution approach to assess the limitations and specific
staining procedures for the applicability of CTFR in P. falciparum invasion phenotyping
assays. Our data show that CTFR effectively labels acceptor erythrocytes and provides a
stable fluorescent intensity at relatively low concentrations. CTFR also yielded a higher
fluorescence intensity relative to DDAO-SE and with a more stable fluorescence intensity
over time. Furthermore, CTFR did not affect merozoites invasion of erythrocytes and was
not toxic to the parasite’s intraerythrocytic development. Additionally, CTFR offers flexibility
in the choice of combinations with several other DNA dyes, which broaden its usage for
P. falciparum erythrocyte invasion assays, considering a wider range of flow cytometers
with various laser settings.
Description
Research Article
Keywords
Cell trace far-red, flow cytometry, Plasmodium falciparum, erythrocyte invasion