Organotypic Brain Slice Cultures Preparation

Two methods are currently used to prepare slice cultures. In the roller-tube technique, pioneered by Gahwiler (1981), the slices are attached to glass coverslips and placed in sealed test tubes on a roller drum in a dry-air incubator. In the membrane or interface technique, pioneered by Stoppini et al. (1991), the slices are placed on semipermeable membranes and grown statically in CO2 incubators. The primary differences between the two techniques are that the rollertube cultures generally become thinner than the membrane cultures but are somewhat more demanding and time consuming to prepare. Detailed culturing procedures can be found elsewhere (e.g., Gahwiler et al., 1998; Thompson and Mason, 2004).

In both methods (Figure 1), brain slices are cut from neonatal animals. The exact age depends on the region of the brain to be cultured. The hippocampus and cortex are typically taken from 5- to 7-day-old pups, whereas the thalamus is reported to survive in culture only when taken from embryonic animals (Molnar and Blakemore, 1991). Slices are cut at a thickness of about 350 mm, as one would for an ordinary acute brain slice experiment. Because of the need to maintain sterility, it is typically easier to use a tissue chopper for the preparation of the slices; however, a vibratome preserves tissue health and integrity better for larger brain sections.

Models of Seizures and Epilepsy

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