Single cell transfection using plasmid decorated AFM probes.

Biochem Biophys Res Commun

Département de pharmacologie, Faculté de Médecine, Université de Sherbrooke, Sherbrooke, Qué., Canada J1H 5N4.

Published: April 2007

AI Article Synopsis

  • Eukaryotic cells are transfected using atomic force microscope tips coated with plasmid DNA for the fluorescent protein EGFP, enabling DNA transfer through the cell membrane.
  • The process involves the tip penetrating the membrane, which can be monitored through cellular deformation, while ensuring minimal damage to the cell.
  • The technique boasts a 30% success rate for transfection and confirms cell survival and growth through epifluorescence microscopy, offering a novel approach to target individual cells.

Article Abstract

Eukaryotic cells were individually transfected using commercially available atomic force microscope tips decorated with plasmidic DNA encoding for the fluorescent protein EGFP. In a typical transfection attempt, the tip is forcibly incorporated into the cell thus allowing for the transfer of the genetic material through the cell membrane. A sharp discontinuity, corresponding to the passage of the tip through the cell membrane can be easily detected when monitoring the cellular deformation as a function of the applied force. In order for the transfection to be successful, the tip must reversibly penetrates the membrane without causing disturbance or damage to the cell. Transfection success rate (30%), cell survival, and growth are confirmed by epifluorescence microscopy. This technique provides an alternative tool to the transfection toolbox, allowing the transfection of specific individual cells with minimal disturbance.

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Source
http://dx.doi.org/10.1016/j.bbrc.2007.01.190DOI Listing

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