FIGURE 1.

Brightfield microscopy of RBCs and RBC aggregates. Microscopic analyses of RBCs and aggregates were performed by a ZOE Fluorescent Cell Imager (Bio-Rad Laboratories, Hercules, CA, United States). Packed RBCs (1 μL) were resuspended in 199 μL of non-aggregating solution (Ringer) and transferred to an ibidi μ-slide. For aggregation analysis, 2.5 μL of packed RBCs were resuspended in 247.5 μL of plasma. The sample was transferred to an ibidi μ-slide and left for 120 s to allow the RBCs to aggregate. (A) control RBCs; (B) aggregates of control RBCs; (C) RBCs from a neuroacanthocytosis patient; (D) RBCs of a neuroacanthocytosis patient after aggregation; (E,F), control RBCs after treatment with orthovanadate before (E) and after aggregation (F); (G,H) control RBCs after treatment with DIDS before (G) and after aggregation (H). The arrows indicate the echinocytes, the large arrowheads point to misshapen RBCs. Treatments were performed as described before (Cluitmans et al., 2012). The length of the bar is 25 μm. Blood was obtained with informed consent and the studies were carried out as described before (De Franceschi et al., 2011; Dinkla et al., 2012; Cluitmans et al., 2015, 2016), in accordance with the CCMO guidelines of the Medical Ethical Committee of the Radboud University Medical Center (file numbers 2007-148, 2013-381, 2018-4421).