Materials and Reagents

1. 12 mm glass coverslips
   
2. 24 well plate
   
3. Pipette tips
   
4. Poly-L-lysine hydrobromide (PLL) (Sigma-Aldrich, catalog number: P1274 )
   
5. Laminin Mouse Protein, Natural (Invitrogen, catalog number: 23017-015 )
   Note: Currently, it is “Thermo Fisher Scientific, Gibco^TM, catalog number: 23017-015”.
   
6. HBSS-CMF (HBSS) (Thermo Fisher Scientific, Gibco^TM, catalog number: 14185-052 )
   
7. Collagenase II (Worthington Biochemical Corporation, catalog number: 4176 )
   
8. Dispase II (Roche Diagnostics, catalog number: 04942078001 )
   
9. Penicillin-streptomycin (Invitrogen, catalog number: 15140122 )
   Note: Currently, it is “Thermo Fisher Scientific, Gibco^TM, catalog number: 15140122”.
   
10. B-27^® Serum-Free Supplement (50x) (Invitrogen, catalog number: 17504-044 )
    Note: Currently, it is “Thermo Fisher Scientific, Gibco^TM, catalog number: 17504-044”.
    
11. Glutamax (Thermo Fisher Scientific, Gibco^TM, catalog number: 35050 )
    
12. Neurobasal-A (Invitrogen, catalog number: 10888-022 )
    Note: Currently, it is “Thermo Fisher Scientific, Gibco^TM, catalog number: 10888-022”.
    
13. 1x Phosphate-buffer saline (PBS) (Roche Diagnostics, catalog number: 11-666-789-001 )
    
14. 4% paraformaldehyde (PFA) (Electron Microscopy Sciences, catalog number: 15710-S )
    
15. Monoclonal Anti-β-Tubulin III antibody produced in mouse (msIgG2b) (Sigma-Aldrich, catalog number: T8660 )
    
16. Goat anti-mouse IgG2b 594 (Invitrogen, catalog number: A21145 )
    Note: Currently, it is “Thermo Fisher Scientific, Novex^TM, catalog number: A21145”.
    

Equipment

1. 37 °C, 5% CO₂ forced air incubator
   
2. Dissecting microscope
   
3. Surgical tools
   
   1. Forceps (Fine Science Tools, catalog number: 11254-20 )
      
   2. Microscissors (Fine Science Tools, catalog number: 15003-08 )
      
   3. Rongeurs (Fine Science Tools, catalog number: 16021-14 )
4. Pipettes
   
5. Hemocytometer
   
6. Microcentrifuge
   

Procedure