Cytometric quantification of singlet oxygen in the human malaria parasite Plasmodium falciparum

The malaria parasite Plasmodium falciparum proliferates within human erythrocytes and is thereby exposed to a variety of reactive oxygen species (ROS) such as hydrogen peroxide, hydroxyl radical, superoxide anion, and highly reactive singlet oxygen (1O2). While most ROS are already well studied in t...

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Bibliographic Details
Published in:Cytometry. Part A 2012-08, Vol.81A (8), p.698-703
Main Authors: Butzloff, Sabine, Groves, Matthew R., Wrenger, Carsten, Müller, Ingrid B.
Format: Article
Language:English
Subjects:
aminophenyl fluorescein (APF)
Animals
Calibration
cercosporin
Flow Cytometry - methods
Humans
Life Cycle Stages - drug effects
Malaria - parasitology
Parasites - drug effects
Parasites - growth & development
Parasites - metabolism
Perylene - analogs & derivatives
Perylene - pharmacology
pixel‐area
Plasmodium falciparum
Plasmodium falciparum - drug effects
Plasmodium falciparum - growth & development
Plasmodium falciparum - metabolism
reactive oxygen species
singlet oxygen
Singlet Oxygen - metabolism
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Summary:The malaria parasite Plasmodium falciparum proliferates within human erythrocytes and is thereby exposed to a variety of reactive oxygen species (ROS) such as hydrogen peroxide, hydroxyl radical, superoxide anion, and highly reactive singlet oxygen (1O2). While most ROS are already well studied in the malaria parasite, singlet oxygen has been neglected to date. In this study we visualized the generation of 1O2 by live cell fluorescence microscopy using 3‐(p‐aminophenyl) fluorescein as an indicator dye. While 1O2 is found restrictively in the parasite, its amount varies during erythrocytic schizogony. Since the photosensitizer cercosporin generates defined amounts of 1O2 we have established a new cytometric method that allows the stage specific quantification of 1O2. Therefore, the parasites were first classified into three main stages according to their respective pixel‐area of 200–600 pixels for rings, 700–1,200 pixels for trophozoites and 1,400–2,500 pixels for schizonts. Interestingly the highest mean concentration of endogenous 1O2 of 0.34 nM is found in the trophozoites stage, followed by 0.20 nM (ring stage) and 0.10 nM (schizont stage) suggesting that 1O2 derives predominantly from the digestion of hemoglobin. © 2012 International Society for Advancement of Cytometry
ISSN:1552-4922
1552-4930
DOI:10.1002/cyto.a.22081