Improvement of Paraffin Sectioning Conditions for the Growing Point of Lilium brownii var.viridulum Baker Bulb
To accurately observe the growth and differentiation process of the growing point of Lilium brownii var.viridulum Baker bulb,this study employed the conventional method to prepare the paraffin sections of the bulbs stored in the refrigerator.The dehydration and transparentizing steps were optimized and the toluidine blue staining method was selected.The conditions for preparing the paraffin sections were improved as follows.First,the scales of a lily bulb were peeled off to expose the growth point,which was then trimmed into a cube of 5 mm×5 mm×5 mm.The cube was fixed in FAA solution for one week,dehydrated with 85%ethanol for 2 h,95%ethanol for 2 h,and 100%ethanol for 3 h,and then transparentized with the mixture of xylene and ethanol(1:1)for 2 h and pure xylene for 3 h.The transparent cube was then immersed in a mixture of paraffin and xylene(volume ratio of 2:3)at 37 ℃ for 2 days,during which a small amount of paraffin was supplemented multiple times.Afterwards,the mixture medium was replaced with pure paraffin in a thermostat incubator at 60 ℃ twice,each time for 4 h.After that,the material was embedded at 60 ℃.The wax block was sliced into pieces with a thickness of 10 μm.After full extension,each slice was put onto a glass slide evenly coated with Herbert gelatin adhesive,and the slide was then heated at 37 ℃ for 5 h.After dewaxing and rehydration,the slide was stained with 1%toluidine blue staining solution diluted by 150 times for 3 min.
Lilium brownii var.viridulum Bakergrowing point of bulbparaffin sectiontechnical improvement
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