Images of honeybush seeds (Cyclopia maculata, Cyclopia genistoides
and Cyclopia subternata) were taken using dissection and light microscopy.
Seedswere softened by soaking inwater overnight (12 h). For fixation
these seeds were placed in a 10% neutral buffered formalin
solution containing 105 g sodium hydroxide, 465 g di-sodium orthophosphate
and 2.5 L of 37% formaldehyde dissolved in 25 L of distilled
water. After formalin fixation, seeds were prepared for wax infiltration.
The penetration of paraffin wax into the seed was found to be impeded
in intact seeds. In order to improve results, the seeds were cut into
halves, the location of the cut depending on the angle of the desired
final section, and folded in permeable paper after fixation, before
being placed into the Sakura Tissue Tek VIP 5 Junior VacuumInfiltration
Processor™ for automatic preparation. The processor carried out dehydration
with ethanol, xylene was used as a clearing agent and paraffin
wax was used as the infiltration media. The following day, the seeds
were embedded in paraffin wax to make wax blocks with the Sakura
Tissue Tek Embedding/Cryo console. Once the paraffin wax had hardened,
the blocks containing the seeds were sectioned to a thickness of
5 μm using a Leica RM 2125 rotary microtome. The prepared sections
were then stained with certified biological stains (Toluidine Blue,
Safranin O orMasson Trichrome) andmounted onto slides. Histological
images and measurements were taken using a Nikon Digital System
DS-L3.