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Biolistic Transfection of Organotypic Brain Slices

he fine processes of the neurons in these slices. Using this method of visualization, even the axons of transfected neurons were visualized for millimeters across the slice, terminating in clearly defined growth cones (Lo et al., 1994).


Discussion
A number of factors affect the success of Biolistic transfection in a given tissue (see Methods and Results above and Sanford et al., 1993 for a more comprehensive review). These parameters must be optimized for each cell and tissue type, and controls must be designed to ensure that bombardment process is not causing cell or tissue damage. For organotypic brain slices, the most important parameter is culture health. If slices are unhealthy for any reason, levels of transfection will be reduced. Once the specific parameters of the Biolistic transfection are optimized for a particular tissue type, the method itself is quite reliable and reproducible. More often than not, if no transfected cells are obtained, it is the health of the cultures that is the problem.

Successful transfection of neuronal cell lines can be achieved with a fairly wide range of Biolistic parameters. The transfection rate of these cells is very reliable, even for co-transfection of multiple plasmids. While transfection rates vary according to the density of cells plated, the amount of gold used in each blast, the efficiency of the expression construct, the exact blasting pressure, the amount of vaccuum pulled, and protecting the cells from the shock of the blast do not seem to be crucial for successful transfection of cell lines.


Summary
We have shown that Biolistic bombardment can be reliably used to transfect neurona
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