Reichert supernova microtome

Anthers were fixed in FAA (10% formaldehyde, 3% acetic acid and 43.5% ethanol) placed under vacuum for 1 h and then keep room temperature. After dehydration in a graded ethanol series and diaphaneity in clearing medium, the material was embedded in Paraffins (from HuaShenPai). Sections (6 μm) were obtained with a Leica Reichert Supernova microtome, placed on glass slides, and stained with a solution of 1%(w/v) toluidine blue O (toluidine blue O 1 g, 95% alcohol 4 mL, 10% acetic acid 10 mL, distilled water 86 mL). Sections examined using a Leica fluorescent compound microscope and Images were captured with a Leica DFC420 camera, and processed with Leica Application Suite software.
The images of whole morphology of WT and mutant were captured using SONY DSC-H50 camera. And the flowers, siliques and seeds morphology were examined using a Leica dissecting microscope.

Anthers with determined stages were first fixed in formalin-acetic acid-alcohol (FAA) under a vacuum for 1 h. After dehydration in a graded ethanol series and diaphaneity in a clearing medium (xylene), the samples were embedded in paraffin (Leica, Weztlar, Germany). Sections (8 μm) were obtained with a Leica Reichert Supernova microtome (Leica, Weztlar, Germany), placed on glass slides, and stained with hematoxylin and eosin (Solarbio, Beijing, China) following manufacturer specifications (Park et al., 1998 (link)). The sections were examined with a fluorescence microscope (Olympus BX43, Tokyo, Honshu, Japan), and images were captured by a CCD attached camera DP73 (Olympus, Tokyo, Japan) with CellSens Standard software.