Materials and Reagents

1. Mice
   
2. Paraformaldehyde (TAAB, catalog number: P001/1 )
   
3. Sucrose
   
4. Glutaraldehyde (Polysciences, catalog number: 111-30-8 )
   
5. Ethanol
   
6. Propylene oxide
   
7. LR White (London resin, medium)
   
8. Hydrogen peroxide
   
9. ImmunoSaver (Nisshin EM, catalog number: 333 )
   
10. Gelatin
    
11. Tween 20
    
12. Uranyl acetate
    
13. Distilled water or Milli-Q water
    
14. Primary antibodies
    
15. 10-nm gold-conjugated secondary antibodies (we purchased from EY laboratories in Tsuru et al., 2013)
    
16. Phosphate buffered saline (PBS) (see Recipes)
    
17. 0.1 M phosphate buffer (pH 7.2) (see Recipes)
    
18. Antigen-retrieval solution (see Recipes)
    

Equipment

1. Peristaltic pump
   
2. Refrigerator
   
3. Freezer
   
4. UV irradiator (Nisshin EM, model: TUV-100 ) with 15 W UV light
   
5. Gold grid (Maxtaform HR25)
   
6. Ultra microtome (Leica Microsystems, model: Ultracut UCT )
   
7. Diamond knife (Diatome AG, catalog number: ultra 45 )
   
8. Incubator
   
9. Microwave (Nisshin EM, catalog number: MWF-2 )
   
10. Electron microscope (Hitachi, model: H-7100 )
    

Procedure