Continuous organelle separation in an insulator-based dielectrophoretic device

Ricardo Ortiz, Domin Koh, Dai Hyun Kim, Mohammad Towshif Rabbani, Cesar Anguaya Velasquez, Mukul Sonker, Edgar A. Arriaga, Alexandra Ros

Research output: Contribution to journalArticlepeer-review

Abstract

Heterogeneity in organelle size has been associated with devastating human maladies such as neurodegenerative diseases or cancer. Therefore, assessing the size-based subpopulation of organelles is imperative to understand the biomolecular foundations of these diseases. Here, we demonstrated a ratchet migration mechanism using insulator-based dielectrophoresis in conjunction with a continuous flow component that allows the size-based separation of submicrometer particles. The ratchet mechanism was realized in a microfluidic device exhibiting an array of insulating posts, tailoring electrokinetic and dielectrophoretic transport. A numerical model was developed to elucidate the particle migration and the size-based separation in various conditions. Experimentally, the size-based separation of a mixture of polystyrene beads (0.28 and 0.87 (Formula presented.) m) was accomplished demonstrating good agreement with the numerical model. Furthermore, the size-based separation of mitochondria was investigated using a mitochondria mixture isolated from HepG2 cells and HepG2 cells carrying the gene Mfn-1 knocked out, indicating distinct size-related migration behavior. With the presented continuous flow separation device, larger amounts of fractionated organelles can be collected in the future allowing access to the biomolecular signature of mitochondria subpopulations differing in size.

Original languageEnglish (US)
Pages (from-to)1283-1296
Number of pages14
JournalElectrophoresis
Volume43
Issue number12
DOIs
StatePublished - Jun 2022

Keywords

  • continuous separation
  • insulator-based dielectrophoresis
  • mitochondria
  • ratchet
  • size-based separation

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry

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