Axiocam

Infected and noninfected (control) roots were collected at 14 to 30 DAI and the galls dissected from roots were submitted to two sectioning procedures.
Fresh galls and uninfected roots were gradually dehydrated in 30, 60, and 90% ethanol and mounted in 3% agarose. Sections of 50 to 300 µm were made with a vibratome (Vibratome 3000 EP; MyNeuroLab). Tissue slices were first mounted on a glass slide for microscopic observations and images were taken with a digital camera (Axiocam; Zeiss). After that, tissues were transferred and attached to a MALDI target plate using an electrical conductive double-sided adhesive tape (3M) and imaged again using the same digital camera coupled to a magnifying glass.
Fresh galls and uninfected roots were mounted in an egg block, as previously described by Sosnowski et al. (2015) , in 10% gelatin or 5% agarose. Subsequently, samples were gradually cooled for 1 h at 4°C and then for 1 h at _ 20°C, and stored overnight at _ 80°C. Sections of 10 to 90 µm were generated at _ 15°C using a cryostat (HYREX C50; Zeiss) in a chamber maintained at _ 35°C. Tissue sections were thaw mounted on ITO glass slides (Bruker Daltonics), dried in a vacuum desiccator, and stored at _ 80°C. Microscopy analyses were performed using brightfield optics and images were obtained with a digital camera (Axiocam; Zeiss).