Begin by cutting sections from intercostal areas alongside the midrib of leaves from Triticum aestivum on a glass plate. Ensure that the length of the cut section runs along the grain of the leaf. Then place the plant material in a syringe with one milliliter of fixation buffer.
Hold the syringe vertically. Remove the air and pump the syringe to create a vacuum holding a finger on the tip. After performing the complete infiltration protocol, as per the resin manufacturer's instructions, cover the cavities of a flat embedding mold with 100%resin, and place the samples on one end of the cavity.
Prepare pencil written paper labels for each sample and place them on the other end of the cavity. Straighten the samples and labels if they moved. Cover the mold with the embedding film and polymerize in an oven as per acrylic resin product guidelines.
Then set the polymerized block with the sample in the ultra microtome specimen holder. Using a rough glass knife, trim the block until the tissue becomes visible and excess resin is eliminated. Next, using a glass or diamond knife cuts semi thin sections transversely.
With a metal inoculation loop, collect the sections from the water surface and place them on a glass slide. Dry the slide on a hot plate to fix the sections onto the glass. Stain the fixed sections with toluidine blue for five seconds.
Semi thin cross sections of Triticum aestivum and Zea mays showed that all leaf tissues were visible and measurable. Zea mays exhibited crans anatomy with mesophyll cells around veins and chloroplast-filled bundle sheaths facilitating rapid metabolite exchange.
