Part I: Isolation of salivary gland sporozoites

Materials and Reagents

1. 15 ml conical centrifugation tube
   
2. Two 10 ml Petri dishes
   
3. Two 27 G needles
   
4. Two 1 ml syringes
   
5. Glass slide
   
6. 1.5 ml plastic reaction tube
   
7. Disposable polypropylene pestles (SP Scienceware - Bel-Art Products - H-B Instrument, catalog number: F19923-0001 )
   
8. Mosquitoes
   
9. Ice
   
10. RPMI (Thermo Fisher Scientific, Gibco^TM, catalog number: 11835063 )
    
11. 70% ethanol
    
12. 1x PBS (Thermo Fisher Scientific, Gibco^TM, catalog number: 18912014 )
    
13. Bovine serum albumin (BSA) (Carl Roth, catalog number: 8076 )
    
14. BSA/RPMI 3% (see Recipes)
    

Equipment

1. Vacuum pump
   
2. Styrofoam box
   
3. Forceps
   
4. Micropipette with disposable tips
   
5. Binocular microscope (ZEISS, model: Stemi 305 or a comparable binocular microscope from any other manufacturer e.g., Nikon, Olympus, Leica)
   
6. Light microscope with phase contrast 40x objective (ZEISS, model: Axio Lab.A1 or a similar device from any other manufacturer e.g., Nikon, Olympus, Leica)
   
7. Hemocytometer
   

Procedure