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Fluorescent Dye Labeling of Erythrocytes and Leukocytes for Studying the Flow Dynamics in Mouse Retinal Circulation
Published on: July 3, 2017
Labeling of Erythrocytes by Porphyrin-Phospholipid
Sunanda Kumar1, Dawei Jiang2, Boyang Sun1
1Department of Biomedical Engineering, State University of New York at Buffalo, NY 14260, USA.
Abstract:
A method is developed for membrane labeling of erythrocytes with porphyrin-phospholipid (PoP). To generate a concentrated PoP solution for labeling human red blood cells (RBCs), various surfactants and solvents are screened to identify conditions that avoid hemolysis, while minimizing non-specific PoP co-precipitation with RBCs in the pellet during centrifugation washes. Cholate, Tween 80 and Tween 40 are identified as useful surfactants for this purpose. When labeled RBCs are mixed with unlabeled ones, substantial non-specific PoP exchange is observed. Egg-yolk lecithin is included in a washing buffer to remove loosely bound PoP and reduce PoP exchange with unlabeled erythrocytes, based on flow cytometry and photodynamic hemolysis assays. Murine RBCs that are labeled with 64Cu-chelated PoP displayed altered biodistribution with longer blood circulation relative to directly administered 64Cu-chelated PoP.
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