油桐花序石蜡切片制作方法改良

【目的】为了获得利用石蜡切片方法观察油桐花芽形成和分化的整个过程的最佳效果,探索针对油桐花序的石蜡切片方法。【方法】以横纵径1 cm左右的油桐花序为实验材料,针对材料处理、脱水、透明、浸蜡、包埋、染色等传统石蜡制片步骤进行改良。【结果】1)保持样品结构不变的情况除去多余苞片、分支以及密实缠绕在小花萼片外的绒毛,使得药液和蜡液更易进入材料内部,避免了由于油桐花序苞片紧实,绒毛极多所导致的脱水、浸蜡不充分等现象;2)改良后的乙醇脱水时间缩短了1 h,并且材料脱水充分,避免了因脱水透明时间过长引起的材料变硬变脆,保证了石蜡可以与材料形成密不可分的整体,切片时能得到完整和连续的蜡带;3)延长浸蜡时间为1 d,保证大体积材料也能浸蜡充分,包埋过程选择在恒温烘箱中进行,保证器械与材料温度一致,避免了两者产生温差导致材料中产生气泡的不良现象,保证材料的完整性。染色后的切片细胞核颜色红亮鲜艳,核质分明,细胞间轮廓清晰,不论是花序的整体形态结构,还是不同组织的细胞结构,均清晰可见。【结论】通过对传统制作石蜡切片方法进行改良有效地解决了油桐花序石蜡切片制作过程中由于其复杂的花序结构及其分泌物质而引起的不良效果以及脱水过度、浸蜡不充分而导致的材料结构不完整、切片易碎等问题,获得了完整连续的蜡带以及结构完整、染色清晰的油桐花序切片。以期望该方法为研究油桐花芽分化及雌雄花发育的细胞学特征提供良好的技术支撑,为快速高效批量制作油桐花序石蜡切片提供了质量保证,同时也为花序结构复杂、外部密披绒毛树种的石蜡切片制作提供参考。

Improvement of paraffin section method on inflorescences of Vernicia fordii

【Objective】In order to obtain the best effect of the whole process of the formation and differentiation of Vernicia fordii flower buds by paraffin sectioning method,and explore the paraffin sectioning method for the Vernicia fordii flower inflorescence.【Methods】The Vernicia fordii inflorescence with a transverse and vertical diameter of about 1 cm was used as an experimental material to improve the traditional paraffin preparation steps such as material processing,dehydration,transparency,waxing,embedding,and dyeing.【Result】1)Keep the sample structure unchanged,remove excess bracts,branches and fluffs that are tightly wrapped around the small calyx,so that the liquid and wax liquid can enter the inside of the material more easily,avoiding the tightness of the twigs of the Vernicia fordii The resulting dehydration,insufficient waxing,etc.;2)the modified ethanol dehydration time is shortened by 1 h,and the material is dehydrated sufficiently,avoiding the hardening and brittleness of the material caused by the excessive dehydration and transparent time,ensuring that the paraffin can be The material forms an inseparable whole,and a complete and continuous wax band is obtained when slicing;3)Extend the waxing time to 1 day,ensure that the bulk material can also be fully waxed,and the embedding process is selected in a constant temperature oven to ensure that the temperature of the device is consistent with the material,avoiding the bad phenomenon that the temperature difference between the two causes bubbles in the material.Guarantee the integrity of the material.The color of the slicednuclei after staining is bright and bright,the nucleoplasm is distinct,and the contours between the cells are clear.The overall morphological structure of the inflorescence or the cell structure of different tissues are clearly visible.【Conclusion】The traditional method of paraffin sectioning was used to effectively solve the adverse effects caused by the complex inflorescence structure and its secretory substances in the process of paraffin sectioning of Vernicia fordii flower inflorescence,as well as the dehydration and insufficient waxing.The material structure is incomplete,the slice is fragile,etc.,and a complete continuous wax band and a structurally intact and well-stained Vernicia fordii inflorescence slice are obtained.This method is expected to provide good technical support for studying the cytological characteristics of flower bud differentiation and male and female flower development,providing quality assurance for rapid and efficient batch production of Vernicia fordii flower inflorescence paraffin sections,as well as complex inflorescence structure and external dense villi.A reference to the production of paraffin sections of tree species.