Tissue Sectioning Procedure

Open valve to CO2 tank and cool stage until completely frozen

Cut straight across cerebellum so brain can stay level and upright

Apply 1% Ethanol to frozen stage as the base

When EtOH is halfway frozen, add a layer of 30% sucrose to the base

When Sucrose is halfway frozen, place brain so that the dorsal side will face towards the blade

Add more sucrose around the tissue and allow to freeze

Manually lower or raise the stage so that the tissue is below where the blade will run

Place blade in its designated space and tighten screws to hold the blade in place

Obtain a brush to handle the tissue and sectioned dish full of distilled H2O to place tissue

Begin cutting tissue into sections of the desired thickness (50 microns) and fully extend cutting arm for automatic adjustments

When done cutting, remove blade, wash with soap and water, then dry. Coat blade in oil and store until next use

For mounting, place one section in dish of water, float to the top and place slide under tissue in water

Using a brush, guide the tissue to its spot on the slide

When done mounting, allow your tissue to dry before staining