Determination Of Electophysiological Properties Of Human Leukocyte Subpopulations Using Lab-On-Chip Device
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Date
2017
Authors
Arif Alsharif, Lina
Journal Title
Journal ISSN
Volume Title
Publisher
Universiti Sains Malaysia
Abstract
The principle of dielectrophoresis (DEP) is based on the polarization and bioparticles movement in applied electric fields. The differentiation of cell population due to DEP force can be plotted in the DEP spectrum. every cell population have specific passing values that will displayed on the DEP spectra, signified change in the directional of frequency-(FDEP) where the applied alternating current (AC) frequency signal increases, then making it a useful tool in differentiating the various cell populations. The current study was carried out to determining the electrophysiological properties of peripheral blood mononuclear cells (PBMC) and monocytes focusing on crossover frequency using lab on chip (LOC) device which utilise microarray dot electrode. PBMC were isolated by density gradient centrifugation technique using ficoll histopaque whereby monocytes were separated by plastic adherence. Then, PBMC and monocytes were subjected to flow cytometry analysis of cell surface markers. Ten l of PBMC and monocytes diluted in DEP medium were added into spacer of LOC device. Subsequently, the alternating current (AC) signal was supplied to observe the dynamic behaviours of PBMC and monocytes in response to the DEP force. The images were captured and analysed using MATLAB software. Images showed that monocytes display weaker p-DEP compared to PBMC. In addition, the sigmoid shape of PBMC DEP spectra were consistent and the cross over frequency occur at 200 kHz. Meanwhile for monocyte DEP spectra, the sigmoid shape were not consistent and the cross over frequency expected to occur between 100 and 200 kHz. In conclusion, the DEP spectra of both PBMC and monocytes is different as expected due to the difference in cellular sizes. The different of DEP
spectra can be used as a criteria in the development of DEP-based characterizing assay in the future.
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Keywords
Determination of electophysiological properties , of human leukocyte subpopulations