Sectioning and Staining

Sectioning

1. Open CO2 valve and cool stage until frozen

2. Cut across cerebellum for brain to stay upright

3. Add 30% sucrose solution when when ethanol is almost forzen

4. When sucrose is almost frozen, place brain so dorsal side faces the blade. 

5. Add sucrose and allow tissue to freeze.

6. Postion stage so the tissue is below where the blade will run.

7. Place blade and tighten screws

8. Obtain a brush and dish with distilled H2O

9. Begin cutting tissue to desired thickness

10. When done, remove blade and clean with soap and water. Coat with oil before storing until next use.

11. Place tissue in distilled water and use a brush to mount it on the slide.

12. Let tissue dry before staining.

Staining

1. First we must rehydrate the tissue

2. Place the tissue mount in H20

3. Then place it in 70% alcohol

4. Then place it in 100% alcohol.

5. Then place it in Clear Safe.

6. Then place the tissue in the same solutions but in the opposite order

7. Then place the slides in the stain.

8. After removing from the stain, place the slides in water.

9. Remove the dirty water and replace with fresh water.

10. Then move it to the 100% alcohol and add water.

11. Then move it to the 70% alcohol

12. Then move it to clear safe and let it sit for a while.

13. Apply paramount to your slides.

14. Place the cover slip on the slide.

15. Do this for the remaining slides and allow to dry.