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Microparticle-enabled single cell multiparameter electronic immunophenotyping for selective electroporation.

Madeline Hoyle1, Josiah Rudge1, Yuvraj Rallipalli1

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Summary
This summary is machine-generated.

This study introduces a microfluidic platform for targeted electroporation (EP) of specific cell types using antibody-labeled microparticles. This method enables precise intracellular delivery to selected cells within a mixed population, advancing cell engineering.

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Area of Science:

  • Biotechnology
  • Cell Biology
  • Microfluidics

Background:

  • Electroporation (EP) is a key non-viral method for intracellular delivery in research and cell therapy.
  • Current bulk EP methods lack the specificity for targeting defined cell subpopulations.
  • Selective delivery is crucial for advanced cell engineering and therapeutic applications.

Purpose of the Study:

  • To develop a microfluidic single-cell platform for targeted electroporation based on surface markers.
  • To enable selective intracellular delivery to specific cell subpopulations within heterogeneous samples.
  • To demonstrate the precision and efficiency of this novel targeted EP approach.

Main Methods:

  • Development of a microparticle-enabled selectively permeabilizing impedance cytometer (ME-SPICy).
  • Utilizing antibody-conjugated microparticles for labeling target cell subpopulations.
  • Implementing multi-parameter impedance sensing for real-time cell discrimination.
  • Applying low-voltage single-cell electroporation based on impedance signatures.

Main Results:

  • ME-SPICy accurately discriminated labeled and non-labeled single cells in real-time.
  • Demonstrated selective electroporation of Jurkat cells and primary cells (PBMCs, T cells).
  • Achieved high purity (up to 98%) and enrichment (>5 fold) in targeted cell populations.
  • Successfully delivered dyes and EGFP mRNA to selected subpopulations.

Conclusions:

  • ME-SPICy provides a precise method for targeted single-cell electroporation.
  • This platform enhances cell engineering by enabling selective intracellular delivery.
  • The technology offers a promising solution to reduce complexity in cell therapy manufacturing.