不同入药部位干药材石蜡制片方法的改良研究

A study on the improvement of the method for paraffin sectioning of dried herbs from different parts of the medicinal plant

石蜡制片作为干药材显微鉴定的关键技术之一,在组织学中应用广泛.根据不同组织构造特点,选取有代表性的皮类、茎类、种子类、根类干药材为主要实验材料,主要从软化、脱水和透明等环节对传统制片方法进行改进,探索各药材专属性石蜡制片方法.结果表明,软化是制作石蜡切片的关键步骤,对不同种类干药材应区别处理.纤维、石细胞富集的皮类生药如厚朴,采用5%氢氟酸水浴浸透软化12 h;木质部纤维化严重的茎类生药如桂枝,采用甘油与氢氧化钠混合溶液软化14 h;内外硬度差异较大的种子类生药如芥子,5%甘油软化6 h;纤维性强和试剂不易渗透的根类生药如黄芪,5%盐酸软化6 h,能提高各干药材横切面的清晰度和完整度,获得组织完整、细胞结构清晰的高质量切片.经改良的石蜡切片技术专属性高,重复性好,避免了常规制片易出现的浸蜡不彻底、组织变硬变脆等问题.研究结果可为制作不同部位石蜡切片提供方法学参考.

Paraffin sectioning is widely used in histology as one of the key techniques for microscopic identification of dried herbs.According to the characteristics of different tissue structures,representative,stem,seed and root dry herbs were selected as the main experimental materials,and the traditional preparation methods were improved mainly from softening,dehydration and transparency to explore the exclusive paraffin preparation methods for each herb.The results showed that softening is the key step in making paraffin sections,and different kinds of dried herbs should be treated differently.Cortex herbs enriched with fiber and stone cells,such as Magnolia officinalis,were softened with 5%hydrofluoric acid in a water bath for 12 h;stems herbs with severe xylem fibrosis,such as Cinnamomum cassia,were softened for 14 h using a mixture of glycerol and sodium hydroxide;seeds of herbs with large differences in internal and external hardness,such as Sinapis alba,were softened with 5%glycerol for 6 h;roots and stems with strong fibers and reagents that were not easy to penetrate,such as Astragalus membranaceus,were sof-tened with 5%hydrochloric acid for 6 h,which could improve the clarity and completeness of the cross section of each dried herb,and obtain high-quality slices with complete tissues and clear cellular structures.The quality of sections with complete or-ganization and clear cellular structure was obtained.The modified paraffin sectioning technique was highly specific and repro-ducible,avoiding the problems of incomplete wax dipping and hardening and brittleness of tissues,which are prone to occur in conventional sectioning.The results of this study could provide a methodological reference for the production of paraffin sections of different parts of the tissue.