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Sample text

Following injection of the construct or constructs, electrodes are used to deliver electric pulses to the area of interest. These pulses induce the transient disruption of the plasma membrane lipids and promote the generation of pores. As a result, the cell permeability increases, allowing the introduction of the plasmids into the progenitor cells, which will then pass on the plasmids to their cellular offspring as they divide. Because DNA is negatively charged at normal pH, the polarity of the electrodes should be considered to direct the DNA into the tissue.

Embryos are handled as gentle as possible. The two embryos nearest the cervix should not be injected. – The mouse is kept hydrated during and after surgery. – Recovery is closely monitored and the animal is checked several times a day after surgery. – If there are any signs of discomfort after 14 h, another injection of buprenorphine is administered. – Pulse parameters are determined in in vitro experiments and then adjusted for the age of the embryos. 1 Age Volts (mV) Pulse length (ms) Interval (ms) No.

30 Isabel Martı´nez-Garay et al. Fig. 4 Clonal lineage analysis of progenitor cells using Cre-activated fluoroproteins. Analysis of clonally derived cells can be carried out by in utero electroporation of STOP-floxed fluorescent labels. 5. (a) Schematic representation of the in utero electroporation site. Fluorescent proteins are expressed only in the cells expressing Cre (IPCs here) but not in radial glia (in black outline). The labels are stably inherited by the neurons. Percentages of labeled cells derived from IPCs in each layer can be described in this manner.

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