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. 2018 Sep 5;8(17):e2990. doi: 10.21769/BioProtoc.2990

Figure 3. Staining of endophytes in perennial ryegrass for CLSM.

Figure 3.

A. Cut tillers close to the root. Tillers can be separated in three parts, the pseudostem (1), the area around the ligule containing the blade meristem (2) and the blade (3). B. Peel the outermost leaf sheath and discard. C. Cut a 2 cm sized piece from the base of the pseudostem (1), the area around the ligule (2) and from the blade (3). D. Cut the pseudostem section (1) in half longitudinally. E. Incubate the leaf samples in 95% ethanol overnight and treat with 10% KOH for 3 h, then wash them three times with PBS. F. Prepare the staining solution. G. Vacuum infiltrate the samples.