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Proceedings Article

Local plasma membrane permeabilization of living cells by nanosecond electric pulses using atomic force microscopy

[+] Author Affiliations
Gary Thompson

Ball Aerospace Inc. (USA) and Clemson Univ. (USA)

Jason A. Payne, Gerald J. Wilmink, Bennett L. Ibey

Air Force Research Lab. (USA)

Caleb C. Roth

General Dynamics Information Technology (USA)

Proc. SPIE 7908, Nanoscale Imaging, Sensing, and Actuation for Biomedical Applications VIII, 79080U (February 15, 2011); doi:10.1117/12.875711
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From Conference Volume 7908

  • Nanoscale Imaging, Sensing, and Actuation for Biomedical Applications VIII
  • Alexander N. Cartwright; Dan V. Nicolau
  • San Francisco, California, USA | January 22, 2011

abstract

Numerous studies provide evidence that nanosecond electric pulses (nsEPs) can trigger the formation of nanopores in the plasma membranes of cells. However, the biophysical mechanism responsible for nanopore formation is not well understood. In this study, we hypothesize that membrane damage induced by nsEPs is primarily dependent on the local molecular composition and mechanical strength of the plasma membrane. To test this hypothesis, we positioned metal-coated, nanoscale cantilever tips using an atomic force microscope (AFM) to deliver nsEPs to localized areas on the surface of the plasma membrane. We conducted computational modeling simulations to verify that the electric field provided by the nsEP is concentrated between the tip and the plasma membrane. The results show that we could effectively deliver nsEPs using the AFM tips at very low voltages. Using scanning electron microscopy we analyzed the tips after applying 10V over 5 seconds duration and found no damage to the tip or loss of platinum coating. As a proof of concept, we applied a 1 and 10V, 5 second pulse to HeLa cells causing large morphological changes. We also applied both a mechanical indention and 600ns electrical pulse stimulus and measured positive propidium ion uptake into the cytoplasm suggesting formation of membrane pores. In future studies, we plan to elucidate the effect that specific, local molecular structures and compositions have on efficacy of electroporation using the newly constructed nano-electrode system.

© (2011) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Citation

Gary Thompson ; Jason A. Payne ; Caleb C. Roth ; Gerald J. Wilmink and Bennett L. Ibey
"Local plasma membrane permeabilization of living cells by nanosecond electric pulses using atomic force microscopy", Proc. SPIE 7908, Nanoscale Imaging, Sensing, and Actuation for Biomedical Applications VIII, 79080U (February 15, 2011); doi:10.1117/12.875711; http://dx.doi.org/10.1117/12.875711


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