Depression slides filled with MS agar medium containing 2% (w/v)

sucrose Individual leaves

Staining solution prepared by mixing 10 mM 2-(N-morpholino) ethanesulfonic acid (MES)-Tris (pH 6.5), 250 mM sucrose, 80 JIM Neutral Red in water. Microscope

CAUTION: MES, Neutral Red, Tris (see Appendix 3) Procedure

1. Prepare depression slides and leaves as described above for FDA staining.

2. Apply the staining solution to the sample and place a coverslip on top. Seal the coverslip with nail polish.

3. Examine the sample in bright field under a compound microscope.

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