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Original
Red Blood Cell Lysis at the Single Cell Level by Using a Mini Electrophoresis Apparatus |
| Chaiyavat Chaiyasut1), Takao Tsuda1), Udompun Khansuwan2) and Siriwan Ong-chai2) |
1)Department of Applied Chemistry, Nagoya Institute of Technology, Gokiso, Showa, Nagoya 466-8555, Japan
2)Department of Biochemistry, Faculty of Medicine, Chiangmai University, Chiangmai, Thailand |
Abstract:
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 (Fenton 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.
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| Keywords: | red blood cell, lysis, single cell, electric field, hydroxyl free radical |
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