1. Place pollen on slide.

2. Incubate in humid chamber (wet Whatman in Tupperware) ~ 30 min.

3. Add drops of FDA (2mg/ml in acetone) to Pollen Growth Media (~/ml) until it becomes turbid.

4. Add 1 drop to pollen.  Incubate 10’

5. Add slide and view under FITC.
 

2X

For 4 Liters:
1360 g sucrose (34%)
16 ml of 1 M CaCl₂ (4mM)
26 ml of 0.5 M Boric Acid (3.25 mM)
pH to 7.5 (~ 2.24 ml of 4 M KOH or 2.24 mM)
 

1X

For 1 Liter:
17% sucrose (170 g)
2 mM CaCl₂ (2 ml of 1 M CaCl2)
1.625 mM Boric Acid (3.25 mL of 0.5 M Boric Acid)
pH to 7.5 with 4 M KOH (1.12 mM; 0.28 ml of 4 M)
bring to volume