1. Insulator-based dielectrophoresis-assisted separation of insulin secretory vesicles
- Author
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Mahta Barekatain, Yameng Liu, Ashley Archambeau, Vadim Cherezov, Scott Fraser, Kate L White, and Mark A Hayes
- Subjects
vesicle isolation ,insulin vesicle ,dielectrophoresis ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
Organelle heterogeneity and inter-organelle contacts within a single cell contribute to the limited sensitivity of current organelle separation techniques, thus hindering organelle subpopulation characterization. Here, we use direct current insulator-based dielectrophoresis (DC-iDEP) as an unbiased separation method and demonstrate its capability by identifying distinct distribution patterns of insulin vesicles from INS-1E insulinoma cells. A multiple voltage DC-iDEP strategy with increased range and sensitivity has been applied, and a differentiation factor (ratio of electrokinetic to dielectrophoretic mobility) has been used to characterize features of insulin vesicle distribution patterns. We observed a significant difference in the distribution pattern of insulin vesicles isolated from glucose-stimulated cells relative to unstimulated cells, in accordance with maturation of vesicles upon glucose stimulation. We interpret the difference in distribution pattern to be indicative of high-resolution separation of vesicle subpopulations. DC-iDEP provides a path for future characterization of subtle biochemical differences of organelle subpopulations within any biological system.
- Published
- 2024
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