Materials and Reagents

1. Arabidopsis plant expressing peroxisome-targeted GFP (Mano et al., 2002)
   
2. Cellulase Onozuka R-10 (Yakult Honsha)
   
3. Macerozyme R-10 (Yakult Honsha)
   
4. MES
   
5. Mannitol
   
6. Enzyme solution (see Recipes)
   
7. Wash solution (see Recipes)
   

Equipment

1. Time tape (Time Med)
   
2. Mending tape (3M)
   
3. Aluminum foil
   
4. 20 ml beaker
   
5. Shaker
   
6. Pasteur pipette
   
7. Test tube
   
8. Swing-out rotor centrifuge machine
   
9. Confocal laser scanning microscope (Zeiss, model: LSM510 META )
   
10. MAS-coated glass slide (super frost) (Matsunami Glass, catalog number: S9441 )
    
11. Cover slip (24 x 60 No.1, 0.12-0.17 mm) (Matsunami Glass, catalog number: C024601 )
    

Software

1. NIH ImageJ software 1.46 for Windows (http://imagej.nih.gov/ij)
   Note: ImageJ is also available on Mac OS X and Linux.
   

Procedure