Anthers fixed in 4% paraformaldehyde were stained in 0.01% (v/v) Renaissance 2200 (Renaissance Chemicals) for 1 h and 10 µg/mL propidium iodide for 1 h. Stained samples were cleared for 48–72 h in ClearSeeAlpha solution44 (link). Samples were embedded in 0.1% agarose gel in a glass capillary. The images were captured using a Lightsheet microscope (Carl Zeiss Z.1) with a 20x Plan Apochromat lens.
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