Part I. Light microscopy

Materials and Reagents

1. 1.5-ml microtubes (Ina-Optika, Bio Bik, catalog number: RC-0150 )
   
2. Mineral wool (Nippon Rockwool, Yasaihana-block50)
   
3. Razor blades (Feather Safety Razor, catalog number: FAS-10 )
   
4. Double-edged razor blades (Feather Safety Razor, catalog number: FA-10 )
   
5. Cell culture dish (Violamo, AS ONE, catalog number: 2-8590-02 )
   
6. Glass microscope slide (Matsunami Glass, catalog number: S1214 )
   
7. Glass microscope cover slip (Trophy, Matsunami Glass, 18 m/m)
   
8. Arabidopsis thaliana ecotype Columbia
   
9. Hydroponic solution (OAT Agrio, OAT House Fertilizer)
   
10. 25% glutaraldehyde (NISSHIN EM, catalog number: 3052 )
    
11. Agar, powder (Wako Pure Chemical Industries, catalog number: 016-11875 )
    
12. Fixing solution (see Recipes)
    
13. 5% agar (see Recipes)
    

Equipment

1. Extra-high-pressure mercury lamp (Nikon Instruments, model: INTENSILIGHT C-HGFI )
   
2. Fluorescence microscope (Nikon Instruments, model: ECLIPSE 80i )
   
3. Stainless-steel pincette (Top well) (AS ONE, catalog number: 5-1076-01 )
   
4. Laboratory glass bottle, screw cap, 100 ml (SCHOTT, DWK Life Sciences, Duran, catalog number: 21 805 24 04 )
   
5. High-pressure steam sterilizer (TOMY SEIKO, model: LSX-500 )
   
6. Microwave (Nisshin EM, model: MWF-2 )
   
7. Refrigerator (Panasonic, model: NR-B52T2-H )
   
8. Micro-slicer (DOSAKA EM, model: DTK-Zero1 )
   
9. Objective lens 20x (Nikon Instruments, model: Plan Fluor 20x/0.50, catalog number: MRH00201 )
   
10. Objective lens 40x (Nikon Instruments, model: Plan Fluor 40x/0.75, catalog number: MRH00401 )
    
11. Objective lens 100x (Nikon Instruments, model: Plan Fluor 100x/1.30 Oil, catalog number: MRH01902 )
    
12. CCD camera (Nikon Instruments, model: DS-Ri1 )
    
13. Condenser (Nikon Instruments, models: D-CUD , D-CUO DIC )
    
14. Autoclave
    

Software

1.Imaging software (Nikon Instruments, model: NIS-Elements)

Procedure