Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Single-cell electroporation.

James L Rae1, Richard A Levis

  • 1Department of Physiology and Biophysics, Mayo Foundation, 200 1st Street SW, Rochester, MN 55905, USA. rae.james@mayo.edu

Pflugers Archiv : European Journal of Physiology
|March 22, 2002
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Altered expression of Ano1 variants in human diabetic gastroparesis.

The Journal of biological chemistry·2011
Same author

Mechanosensitivity of Nav1.5, a voltage-sensitive sodium channel.

The Journal of physiology·2010
Same author

Sodium channel mutation in irritable bowel syndrome: evidence for an ion channelopathy.

American journal of physiology. Gastrointestinal and liver physiology·2008
Same author

The lens: local transport and global transparency.

Experimental eye research·2004
Same author

Properties of single voltage-gated proton channels in human eosinophils estimated by noise analysis and by direct measurement.

The Journal of general physiology·2003
Same author

alpha(1C) (Ca(V)1.2) L-type calcium channel mediates mechanosensitive calcium regulation.

American journal of physiology. Cell physiology·2002

This study introduces a novel electroporation technique using modified patch-clamp methods to efficiently insert genes and compounds into single cells. This method achieves high transfection rates with minimal cell damage, offering a versatile alternative for cellular delivery.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biotechnology

Background:

  • Traditional methods for intracellular delivery, such as pressure injection and iontophoresis, have limitations.
  • Efficient and targeted delivery of genetic material and compounds into single cells remains a challenge.

Purpose of the Study:

  • To develop and validate a modified patch-clamp technique for routine gene and compound delivery into single cells via electroporation.
  • To optimize electroporation parameters for high transfection efficiency and minimal cell damage.

Main Methods:

  • Utilized a modified patch-clamp setup with a small-tipped microelectrode for cell indentation.
  • Applied low voltages (2-10 V) via rectangular pulses (20 µs to >300 ms) or DC to 5 kHz frequencies.
  • Focused on optimizing the total duration of voltage application for effective electroporation.

Related Experiment Videos

Main Results:

  • Achieved routine transfection rates exceeding 80% with optimal parameters.
  • Demonstrated that membrane dielectric breakdown creates pores for compound entry.
  • Showed that larger genes required higher voltages for successful delivery.

Conclusions:

  • Modified patch-clamp electroporation is an effective alternative for delivering genes, drugs, and other compounds into cells.
  • The technique allows precise delivery from various cell surface locations with minimal cellular damage due to small electrode tips.
  • This method offers improved efficiency and reduced invasiveness compared to existing intracellular delivery techniques.