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Water Bath Mounting Techniques

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WATER BATH MOUNTING TECHNIQUES

  1. The information below is general technique.
  2. The ribbon is separated from the knife edge with a pick or tweezers and pulled across the surface of a warm water bath. The temperature of the water should be 8-10°C below the melting point of the wax used in the block (we set ours at 42°C) Wrinkles in the section can be removed along with small air bubbles trapped beneath the wax, by careful teasing from below with a metal probe.
  3. Sections can be separated whilst floating on the water with gentle pressure from the tips of forceps. (They will also separate easily when the edge of the slide is lined up with the seam as you lift it out of the water).
  4. Sections are collected by holding the slide mostly beneath the surface of the water. The slide is carefully moved against the outer edge of the section. The slide is lifted vertically (@ 900) from the water. The water will sheet out from under the section as the slide is lifted. If water becomes trapped by the outer edges of the paraffin, you can gently remove it by laying the slide on a firm surface, placing a damp fine filter paper or lens over the section, and pressing lightly across the section from top to bottom. This will force any remaining water from under the tissue and insure complete contact between the section and the slide surface. Carefully lift the damp paper from the slide. This technique will also work for sections mounted by the hot plate method.
  5. Slides are air dried in a vertical position for a minimum of 10 minutes to   ensure the section is firmly attached. The slides are then dried in an oven set at 60°C for a specified length of time depending on the staining protocol to be used.