Cutting and Mounting Sections

1. Cut 7 to 10-pm sections as ribbons on a rotary microtome using a disposable blade.

2. Ribbons are placed on a pool of 20% (v/v) ethanol in DEPC-treated water on a slide, and cut into smaller strips with a clean razor blade if required.

3. The sections are floated out onto DEPC-treated water in a clean section bath at 45°C until creases in them disappear.

4. Sections are then collected onto TESPA-treated slides, and dried on a 48°C hot plate for 2 h or overnight at 37°C. It is useful to collect at least two adjacent sections per slide; this aids the subsequent identification of hybridizing tissues, since these should give identical results on both sections.

5. Store slides desiccated at 4°C such that they are free from moisture and sections still give acceptable results months after being cut. Before use, allow slides to warm to room temperature. If some slides are to be returned to storage, place them in the 37°C oven for 30 min prior to storage to reduce the moisture and store sealed with fresh desiccant.

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