Materials and Reagents

1. Superfrost plus Microscope slide (Thermo Fisher Scientific, catalog number: 12-550-17 )
   
2. Cover Glass (Thermo Fisher Scientific, catalog number: 12-545-88 )
   
3. Tissue sample
   
4. Fluoro-Jade B (Merck Millipore Corporation, catalog number: AG310 )
   
5. Paraformaldehyde (Electron Microscopy Science, catalog number: 19210 )
   
6. Potassium permanganate (KMnO₄) (Sigma-Aldrich, catalog number: 223468 )
   
7. DAPI (Life Technologies, catalog number: D3571 )
   Note: Currently, it is “Thermo Fisher Scientific, Molecular Probes^TM, catalog number: D3571”.
   
8. Glacial Acetic acid (CH₃CO₂H) (Sigma-Aldrich, catalog number: A9967 )
   
9. Ethanol
   
10. Xylene (Sigma-Aldrich, catalog number: 534056 )
    
11. Sodium hydroxide (NaOH) (Sigma-Aldrich, catalog number: S8045 )
    
12. Sodium tetraborate decahydrate (Sigma-Aldrich, catalogue number: B9876 )
    
13. Sodium chloride (NaCl) (Sigma-Aldrich, catalog number: S3014 )
    
14. potassium phosphate dibasic (K₂HPO₄) (Sigma-Aldrich, catalog number: P3786 )
    
15. potassium phosphate monobasic (KH₂PO₄) (Sigma-Aldrich, catalog number: P9791 )
    
16. 4% paraformaldehyde (see Recipes)
    
17. KPBS (see Recipes)
    
18. 0.2% Fluoro-Jade (see Recipes)
    
19. Fluoro-Jade solution (see Recipes)
    
20. 0.2% DAPI (see Recipes)
    

Equipment

1. Vacuum Desiccators (Thermo Fisher Scientific, catalog number: 08-642-5 )
   
2. Tissue-Tek slide staining set (Electron Microscopy Science, catalog number: 62540-01 )
   
3. 24 slide holder (Electron Microscopy Science, catalog number: 62543-06 )
   
4. Orbital shaker
   
5. Timer
   
6. Slide Warmer
   
7. DPX mounting medium (a mixture of the polystyrene distyrene and the plasticizer dibutylphthalate) (Electron Microscopy Science, catalog number: 13512 )
   

Procedure