Implantation of Dkk-1-soaked Beads into the Neural Tube of Chicken Embryos

Materials and Reagents

1. Fertilized chick (Gallus gallus)
   
2. Recombinant mouse Dkk-1 (R&D Systems, catalog number: 1765-DK )
   
3. Heparin Acrylic beads (Sigma-Aldrich, catalog number: H5263 )
   
4. Bovine Serum Albumine (BSA) (Sigma-Aldrich, catalog number: A2153 )
   
5. Pelikan Drawing Ink Black
   
6. Silikon Peroxid (IDEX Health & Science, catalog number: SC0083ST )
   
7. Tungsten wire (0.380 mm, 10 FT) (World Precision Instruments, catalog number: TGW1510 )
   
8. Grid with concentric circles (NE42-21 mm) (Graticules LTD, Tonbridge Kent)
   
9. Penicillin-Streptomycin (10,000 U/ml) (Gibco, catalog number: 15140-122 )
   
10. NaCl (Sigma-Aldrich, catalog number: S-3014 )
    
11. KCl (Sigma-Aldrich, catalog number: P-9541 )
    
12. CaCl₂^.2H₂O (Prolabo, catalog number: 22317297 )
    
13. NaH₂PO₄^.2H₂O (Scharlab, catalog number: SO0334 )
    
14. MgCl₂^.6H₂O (Prolabo, catalog number: 27778293 )
    
15. D(+) glucose anhydre (Prolabo, catalog number: 24370294 )
    
16. 10x Phosphate-buffer saline (PBS) (see Recipes)
    
17. 4% Paraformaldehyde (PFA) from 20% PFA (see Recipes)
    
18. PBS/0.1% BSA (see Recipes)
    
19. Tyrodes buffer (see Recipes)
    
20. 0.5% Bicarbonate (see Recipes)
    
21. 0.5% Glucose (see Recipes)
    
22. Tyrodes supplemented (see Recipes)
    

Equipment

1. 37 °C, 5% CO₂ forced-air incubator (Novital, model: Covatutto 120-4V )
   
2. Dissecting microscope (Leica Microsystems, model: MS5 )
   
3. Butane burner (Butsir)
   
4. Glass micropipet (homemade)
   
5. Pasteur pipets (Deltalab, catalog number: 200001 )
   
6. Silicon tube (IDEX Health & Science, catalog number: SC0083ST )
   

Procedure