First open valve to CO2 tank and cool stage until it is completely frozen

Next, cut straight across the cerebellum so the brain can stay level and upright

Then, apply 1% ethanol to the frozen stage as the base

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

When the sucrose if halfway frozen, place the brain so that the dorsal side will face towards the blade

Add more sucrose around the tissue and allow it to freeze

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

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

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

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

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

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

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

When done mounting, allow your tissue to dry before staining

The slides are then finished!

Take 6 containers and place the 6th in the back

Add distilled water to the first container

Add 70% alcohol to the second container

Add absolute alcohol to the third container

Add clear safe to the fourth container

Add cresyl violet stain to the fifth container which is placed to the left of all the other containers

Take the slides and make sure the side that has the glass is facing outwards

Place the slide in the first container containing distilled water

Place each slide in the slots in the glass container and leave them there for about two minutes before moving on to the next step

Take each slide out of the distilled water and place them in the 70% alcohol

Leave them in for 2-4 minutes (closer to four minutes) to ensure all the tissue is dehydrated

Move the slides into the absolute alcohol and keep them there for about five minutes

Then move the slides into the clear safe

• if you see a smoke like appearance in the water, move them back into the alcohol

• if there isn't a smoke like appearance, keep the slides in the clear safe for about five minutes or until the tissue becomes clear

• steps 5-8 are a reversal of the steps listed above

• put them in the alcohol for 4 minutes and then into the 70% alcohol for another 4 minutes

• place them in the distilled water for 2 minutes and then place the slides in the cresyl stain container

• leave the slides in the stain for about 20 minutes although the literature says 10

• add acid alcohol to the sixth container that was placed in the back previously

• then place this container in between the water and the 70% alcohol containers

• move the slides out of the stain and into the water

• once the water changes colors, dump it and refill with distilled water

• moce the slides into the acid alcohol and repeat the dumping and refilling step

• move the slides into the 70% alcohol and leave this here for as long as necessary

• move the slides into the absolute alcohol and let them sit for a little bit longer

• move the slides to the clear safe for a while

• next, prepare the cover slips

• grip the edges of the coveer slips to separate them

• lay the cover slips in the bin so that they stick out to be easily grabbed

• open the paramount bottle with gloves

• place a one-sided q-tip into the paramount bottle

• grab one slide from the clear safe and make sure the side with the brains is facing upwards

• hold the slide at an angle facing away from you

• take the q-tip and place the paramount on the bottom portion of the slide that is facing away from you

• take a cover slip and hold it perpendicular to the slide

• place the cover slip over the slide

• tap it to ensure each side is even

• then you are done w the staining process

• leave the slides out ona flat surface to dry