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Spatiotemporally controlled single cell sonoporation.

Zhenzhen Fan1, Haiyan Liu, Michael Mayer

  • 1Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109, USA.

Proceedings of the National Academy of Sciences of the United States of America
|September 27, 2012
PubMed
Summary

This study demonstrates precise control over ultrasound-induced cell membrane disruption (sonoporation) at the single-cell level. Researchers precisely delivered molecules and measured cellular transport and drug efflux with high repeatability.

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Area of Science:

  • Biophysics
  • Cell Biology
  • Biotechnology

Background:

  • Ultrasound-mediated sonoporation offers potential for targeted cellular delivery.
  • Precise control and quantification of sonoporation at the single-cell level remain challenging.
  • Understanding pore dynamics and intracellular transport is crucial for therapeutic applications.

Purpose of the Study:

  • To develop and validate methods for precise control and quantification of single-cell sonoporation.
  • To investigate the spatiotemporal dynamics of sonoporation-induced membrane pores.
  • To measure intracellular transport and drug efflux in individual cells.

Main Methods:

  • Utilized ultrasound-excited microbubbles targeted to cell membranes for controlled sonoporation.
  • Employed whole-cell patch clamp recording and fluorescence microscopy for time-resolved measurements.
  • Quantified pore size, resealing rates, intracellular diffusion coefficients, and drug efflux.

Main Results:

  • Achieved high repeatability in spatially and temporally controlled sonoporation.
  • Quantified single-cell pore dynamics and intracellular diffusion of RNA/DNA.
  • Demonstrated precise delivery of molecules, including a drug substrate, with subcellular accuracy.
  • Successfully measured calcein efflux mediated by multidrug resistance protein 1 (MRP1) in individual cells.

Conclusions:

  • Developed a robust platform for controlled single-cell sonoporation and molecular delivery.
  • Enabled precise quantification of membrane pore dynamics and intracellular transport.
  • Provided a method for measuring drug efflux in individual cells, facilitating drug resistance studies.