Establishment of Cryosectioning Method and Histochemical Localization Analysis of Volatile Oil and Coumarin Components of Ferula ferulaeoides
10.13422/j.cnki.syfjx.20211860
- VernacularTitle:多伞阿魏的冰冻切片方法建立及挥发油、香豆素类成分组织化学定位分析
- Author:
Yuanyuan ZHAO
1
;
Ying MA
1
;
Shuangshuang LIU
1
;
Mengmeng LIU
1
;
Xin LU
1
;
Jiaqi YAO
1
;
Siwei ZHANG
1
;
Yun ZHU
1
Author Information
1. Key Laboratory of Xinjiang Phytomedicine Resource and Utilization，Ministry of Education， School of Pharmacy，Shihezi University，Shihezi 832000，China
- Publication Type:Journal Article
- Keywords:
Ferula ferulaeoides;
frozen section;
volatile oil;
coumarin;
resin canal;
sucrose
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2022;28(13):161-166
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo establish the frozen section method of Ferula ferulaeoides， and to study the histochemical localization of volatile oil and coumarins in different organs of F. ferulaeoides. MethodThe roots， stems， petioles and leaves of F. ferulaeoides were used as materials to investigate the concentration of sucrose protectant， liquid nitrogen flash-freezing time， embedding conditions， section thickness， freezing temperature and time and post-treatment methods， the most suitable section conditions were screened by comparing the integrity， microscopic effect， elongation and clarity of frozen sections. Sudan Ⅳ staining method and fluorescence microscopy were used to locate the volatile oil and coumarins of F. ferulaeoides. ResultThe optimal conditions for frozen sections of the roots， stems， petioles and leaves of F. ferulaeoides were as follows：10%， 15% and 20% gradient sucrose as the protectant for roots， 10%， 20% and 30% gradient sucrose as the protectant for stems and petioles， 20%， 25% and 30% gradient sucrose as the protectant for leaves， glue-water （2∶1） as the embedding agent， quick-freeze in liquid nitrogen for 20 s， warmed up at -25 ℃ for 30 min， sliced at -20 ℃ with the thickness of 25 μm， rinsed with the same concentration of sucrose solution （gradient sucrose solution selected the last concentration）， and the slices placed on the ice pack for a period of time and stored at room temperature. Among them， the concentration of sucrose protectant was the most important factor. The results of histochemical localization showed that volatile oil and coumarins in four organs of F. ferulaeoides were mainly distributed in resin canal. ConclusionFrozen section of F. ferulaeoides is established for the first time with high rate of slicing and simplified steps， its volatile oil and coumarins are mainly accumulated in resin canal.
