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- Title
Insulator-based dielectrophoresis-assisted separation of insulin secretory vesicles.
- Authors
Barekatain, Mahta; Yameng Liu; Archambeau, Ashley; Cherezov, Vadim; Fraser, Scott; White, Kate L.; Hayes, Mark A.
- 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 highresolution separation of vesicle subpopulations. DC-iDEP provides a path for future characterization of subtle biochemical differences of organelle subpopulations within any biological system.
- Subjects
DIELECTROPHORESIS; SECRETORY granules; INSULIN; PROTEIN fractionation; BIOLOGICAL systems; INSULINOMA
- Publication
eLife, 2024, p1
- ISSN
2050-084X
- Publication type
Article
- DOI
10.7554/eLife.74989