Rm2125 microtome

The tested specimens, including the upper leaf developed on the third branch from the base of the main stem of chili plants, were selected during the second growing season of 2022, 10 days after the third foliar application. A sample of approximately 1 cm of the specimens was taken from each treatment, killed, and fixed in FAA solution (5 mL of glacial acetic, 10 mL of formalin, 35 mL of water, and 50 mL of ethyl alcohol 70%), for 48 h at least. The chosen sample was washed in ethyl alcohol at 30%, dehydrated in a normal ethanol and butyl alcohol chain, buried in paraffin wax with a melting point of 56 °C, sliced to a thickness of 15 μm, stained with crystal violet and erythrosin, clarified in xylene, and mounted in Canada balsam. Transverse sections were obtained using a Leica Microtome RM 2125, micrographed, and measured using a Leica Light Image Analysis System DM 750 at the Research Park (CURP), Faculty of Agriculture, Cairo University. The following parameters were recorded: the thickness of the midvein (μm), lamina (μm), palisade tissue (μm), spongy tissue (μm), upper and lower epidermis (μm), bundles dimension (μm), and mean vessels diameter (μm).

Tested specimens included the blade of the terminal leaflet of the fourth compound leaf developed on the main stem of tomato plants under low-temperature conditions, were taken throughout the second growing season of 2021/2022, 60 days after sowing. Approximately 1.0 cm of the specimens were killed and fixed in FAA solution (5 mL glacial acetic, 10 mL formalin, 35 mL water, and 50 mL ethyl alcohol 70%) for at least 48 h. The selected materials were washed in 30% ethyl alcohol, dehydrated in a normal ethanol and butyl alcohol series, embedded in paraffin wax with a melting point of 56 °C, sectioned to a thickness of 15 μm stained with crystal violet erythrosin, cleared in xylene, and mounted in Canada balsam in accordance with [70 ]. Transverse sections were completed with a Leica Microtome RM 2125 and then micrographed and measured using a Leica Light Image Analysis System DM 750 at the Faculty of Agriculture, Cairo University-Research Park (CURP). The following parameters were recorded: thickness of the midvein (μm), lamina (μm), palisade tissue (μm), spongy tissue (μm), upper and lower epidermis, bundle dimension (length and width) (μm), and mean vessels diameter (μm).

Histology was performed using an established protocol (Sullivan-Brown et al., 2011 (link)) with the following modifications. Whole adult zebrafish were fixed in Dietrich's fixative (ZIRC Health Services Sample/Specimen Preparation) (30:10:2:58 ratio of 95% ethanol:formalin:glacial acetic acid:distilled water) for 1-2 weeks, followed by dehydration (20 min incubation step in 50% ethanol in PBS, followed by a 20 min incubation step in 75% ethanol in PBS and two 20 min incubation steps in 100% ethanol, all at room temperature), JB-4 infiltration and embedding. Embedding was performed overnight under static vacuum at 4°C. Then, 4-µm sections were collected on a Leica microtome (RM2125) with 8 mm glass blades and stained with H&E based on a previously published protocol (Sullivan-Brown et al., 2011 (link)).