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Quantification and Size-profiling of Extracellular Vesicles Using Tunable Resistive Pulse Sensing
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Resistive-Pulse Sensing Inside Single Living Cells.

Rongrong Pan1,2, Keke Hu1,3, Rui Jia1,3

  • 1Department of Chemistry and Biochemistry, Queens College-CUNY, Flushing, New York 11367, United States.

Journal of the American Chemical Society
|March 3, 2020
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Summary
This summary is machine-generated.

Researchers developed localized resistive-pulse sensing to detect nanoscale biological entities within living cells. This technique enables the in situ analysis of cellular vesicles and nanoparticles, advancing intracellular sensing capabilities.

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Area of Science:

  • Biophysics
  • Nanotechnology
  • Cell Biology

Background:

  • Resistive-pulse sensing detects nanoscale entities in bulk solutions.
  • Localized sensing within living cells has not been previously reported.
  • This technique is crucial for understanding cellular processes and disease mechanisms.

Purpose of the Study:

  • To develop and demonstrate localized resistive-pulse sensing within living cells.
  • To detect and characterize intracellular vesicles and nanoparticles.
  • To perform electrochemical analysis of reactive species within single vesicles.

Main Methods:

  • Utilized a nanopipette as a scanning ion conductance microscopy (SICM) tip for resistive-pulse experiments.
  • Monitored cell membrane penetration by characteristic ion current changes.
  • Performed in situ detection of cellular vesicles and nanoparticles within macrophages.
  • Conducted electrochemical resistive-pulse experiments using a conductive carbon nanopipette.

Main Results:

  • Successfully detected cellular vesicles (phagosomes, lysosomes, phagolysosomes) inside RAW 264.7 macrophages.
  • Achieved selective detection of 10 nm gold nanoparticles within the macrophage cytoplasm.
  • Demonstrated in situ detection of extracellular vesicles from metastatic human breast cells (MDA-MB-231).
  • Measured reactive oxygen and nitrogen species (ROS/RNS) within single vesicles.

Conclusions:

  • Localized resistive-pulse sensing is feasible for intracellular and in situ analysis.
  • This method allows for the detection and characterization of nanoscale biological entities within their native cellular environment.
  • The technique offers potential for advanced diagnostics and understanding of cellular functions and diseases.