Red Blood Cell Lysis at the Single Cell Level by Using a Mini Electrophoresis Apparatus

Observation of the lysis phenomenon of human erythrocyte at the single cell level under a mini electrophoresis apparatus was studied. The lysis-inducing reagent (hydrogen peroxide in saline) was introduced to the uptake ferrous red blood cells at the end tip of the rectangular capillary by electroosmotic flow, generating hydroxyl free radical (Fen ton reaction). Red blood cell exposed to hydroxyl free radical underwent progressive lysis, determined by observation through a microscope-CCD camera recording system. The erythrocyte lysis time of several single red blood cells varied between 450 sec and 510 sec. Distinct lysis behavior was delineated by monitoring the apparent light density of each single erythrocyte through time. The time dependence apparent light density curve of single red blood cell was used to assess the lysis phenomenon. It was found that the rate of red blood cell alteration dramatically increased after some initial period leading to eventually complete lysis. The half-lives of the initial step and the second step were estimated as kinetic parameters. The proposed technique stabilized the lysis mechanism by minimizing the depletion effect of the enveloped solvent layer of red blood cell. Moreover, electric field strength provided more homogeneity of the enveloped solvent layer and enabled the erythrocyte to maintain at the observed position.