Materials and Reagents

1. Mouse pancreas
   
2. Ethanol (100%, 95%, 85%, 70%, 50%)
   
3. 0.85% NaCl
   
4. ddH₂O
   
5. 99.5% Xylene (Sigma-Aldrich, catalog number: 534056 )
   
6. 10% Formalin solution (neutral buffered) (Sigma-Aldrich, catalog number: HT501128 )
   
7. Bovine Serum Albumin (BSA) (Sigma-Aldrich, catalog number: A7906 )
   
8. Tween 20 (Sigma-Aldrich, catalog number: P9416 )
   
9. Triton-X 100 (Sigma-Aldrich, catalog number: T8787 )
   
10. Guinea pig anti-insulin antibody (Millipore, catalog number: 4011-01F )
    
11. Rabbit anti-glucagon antibody (Millipore, catalog number: 4030-01F )
    
12. Mouse anti-EGFP antibody (Clonetech, catalog number: 632381 )
    
13. Texas Red conjugated donkey anti-guinea pig antibody (Jackson ImmunoResearch Laboratories, catalog number: 106-075-003 )
    
14. Alexa 350 conjugated goat anti-rabbit antibody (Life Technologies, Invitrogen^TM, catalog number: A-21068 )
    
15. FITC conjugated goat anti-mouse antibody (Jackson ImmunoResearch Laboratories, catalog number: 715-095-166 )
    
16. ProLong Gold Antifade Reagent (Life Technologies, Invitrogen^TM, catalog number: P36935 )
    
17. 10x PBS (Gibco^®, catalog number: 70011-044 )
    
18. Blocking solution (see Recipes)
    

Equipment

1. Glass jar
   
2. Humidified chamber
   
3. Confocal microscope
   
4. Tissue cassette
   

Procedure