By Jac A. Nickoloff
Harvard college, Boston, Massachusetts. handbook of protocols within the electroporation of proteins and DNA into fish, insect, and mammalian cells. For researchers. Plastic comb spiral binding. sixty five participants, 28 U.S.
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Additional resources for Animal Cell Electroporation and Electrofusion Protocols (Methods in Molecular Biology)
For fusion, a continuous flow is not desirable, because fused cells need to be undisturbed for a period of time to round off and complete the fusion process. In this case, a pulsating (stop and go) flowthrough system would be appropriate. 4. Generators The relationship between the electrical parameters in a generator and the parameters actually delivered to the chambers is important because substantial differences can exist. Following this discussion, different types of generators are described.
Clearly there is merit in using lower applied field strength, and increasing the duration and number of pulses to achieve the required ET value. Protocols for electroporation continue to develop as cases demand. The present knowledge of electroporation mechanisms is capable of guiding rational design of new protocols. Instead of describing a cumulation of successful protocols, this chapter gives the basics to develop one’s own protocols. It is hoped that the above analysis will take some guesswork out of applying electrotransfection in new situations.
3 e cm for nonionic solutions, such as mannitol. The resulting current density j is: j = E/r (Alcm2) (2) The current produced results in heating of the medium. Saline solutions with a low value of r experience severe heating effects as compared to nonionic solutions for the same electric field and pulse length. The temperature rise AT (“C) can influence the permeabilization mechanism, or lead to excessive heating and evaporation of the medium. 2 r, where t is the pulse length in s. ). Having defined the parameters at the location of the cells, we can relate them to the electrical parameters at the chamber electrodes: Electrode voltage: V = E .