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Fig. 2 | Malaria Journal

Fig. 2

From: A simple monochromatic flow cytometric assay for assessment of intraerythrocytic development of Plasmodium falciparum

Fig. 2

Optimization of VSG staining of P. falciparum-infected erythrocytes. a Gate setting for flow cytometric analysis. Non-single cells were excluded by gating according to FSC-H, FSC-W, SSC-H, and SSC-W. b Histograms show fluorescence intensity of VSG+ cells (green) excited by 488-nm, 633-nm, and 375-nm lasers. To read the emitted fluorescent signal, detectors of FITC (500–560 nm), PE (543–627 nm), PE-Texas Red (593–639 nm), PerCP-Cy5-5 (655–735 nm), and PE-Cy7 (720–840 nm) were used for 488-nm activating laser; detectors of APC (640–680 nm), A700 (685–775 nm), and APC-Cy7 (720–840) were used for 561-nm activating laser; and, detectors of BV421 (400–500 nm), BV510 (500–560 nm), and BV605 (590–630 nm) were used for 445-nm activating laser. Histogram of sample not stained with VSG was set as VSG negative (shown in magenta). c Representative flow cytometric profiles of samples stained with VSG at 0.5, 1, 2, 5, 10, and 20 µg/mL relative to the 10,000x concentration (20 mg/mL) of the commercial version. Overlaid histogram of VSG+ cells obtained from staining with different concentrations of VSG is shown on the left side of flow cytometric images. d Representative images of Giemsa-stained erythrocytes in VSG+ fraction acquired using an objective lens at 100×. Cells were sorted from the sample stained with 10 µg/mL of VSG. Scale bars: 10 µm. FSC-A, forward scatter area; FSC-H, forward scatter height; FSC-W, forward scatter width; SSC-W, side scatter width; SSC-H, side scatter height; DIC, differential interference contrast; VSG, ViSafe Green

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