Ni e light microscope

Paraffin sections from each sample were first cut onto slides, dewaxed in xylene, dehydrated in graded ethanol, and microwaved at high intensity in citrate buffer (pH 6.0). Then, the sections were treated with 3% H₂O₂ in PBS for 10–30 min to block endogenous peroxidase, followed by blocking with 2–5% normal bovine serum albumin in PBS (pH 7.4) for 1 h. For the endosperm nuclei or root cells/nuclei, they were first spread onto the slides directly and then allowed to dry at room temperature. Next, all sections were incubated with primary antibodies (anti-HAG704 or anti-ACLA2, homemade) diluted 1:100 and the secondary antibody (Invitrogen, A-11011) diluted 1:200 in PBST (pH 7.4, PBS with 0.1% Tween 20) at room temperature for 2 h, respectively. After three washes with PBST, the sections were stained with DAPI and viewed with a Nikon Ni-E light microscope equipped with a CCD camera.

Rice seeds were collected and fixed in FAA (4% formaldehyde, 10% acetic acid, and 50% ethanol) before being dehydrated in a series of graded concentrations of ethanol. These seeds were polymerized at 37 °C for three days before being embedded in paraffin. The embedded samples were first cut into 10 µM thick sections and counterstained with periodic acid-Schiff reagent and toluidine blue before being examined using a Nikon Ni-E light microscope with a CCD camera.

Liver and fat tissues were fixed with 10% formalin for at least 24 h. Samples were sliced into 5 μm sections after embedding in paraffin and then stained with H&E. Digital images were captured with a Nikon NI-E light microscope (Nikon, Tokyo, Japan).