( air. After the tissue is placed in the ...)Biolistic Transfection of Organotypic Brain Slices

air. After the tissue is placed in the chamber, the chamber vacuum is drawn down to 23" Hg. The access valve is then opened and the chamber is refilled with helium from a second tank. Subsequently, the chamber vacuum is reduced to 25" Hg and held until the particles are accelerated into the tissue. Reducing the density of the overlying gas both decreases the drag on the gold particles and lessens the force of the shock wave.

Brain Slice Results
Following Biolistic transfection, neurons and glia expressed β-galactosidase at high levels as detected by X-gal histochemistry (see Lo et al., 1994). Figure 3 shows a low-power view of a slice to illustrate the number of transfected cells in a slicegenerally several hundred cells are transfected per slice, located throughout the entire depth of the cortical plate. Expression can be detected as early as 8 hours post-transfection and increases to fill all of the small processes of both the dendrites and axons of the neurons by 36 hours (Figure 4). Expression lasts for at least 7 days, although the end-point of this transient transfection was not determined. In all transfected cells, a single gold particle can be seen in the nucleus (Figure 4).

Because fine dendritic processes cannot be resolved using X-gal staining (its chromogenic reaction product diffuses some distance away from the enzyme), we visualized these transfected neurons using immunostaining to locate β-galactosidase (Lo et al., 1994; primary antibody obtained from 5 Prime-3 Prime; secondary rhodamine-conjugated goat anti-rabbit antibody from Boehringer-Mannheim). Figure 4 illustrates how effectively immunostaining increases the resolution of t
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