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Establishment of Cryosectioning Method and Histochemical Localization Analysis of Volatile Oil and Coumarin Components of Ferula ferulaeoides / 中国实验方剂学杂志
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 161-166, 2022.
Article in Chinese | WPRIM | ID: wpr-940672
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. Sudanstaining 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.

Full text: Available Index: WPRIM (Western Pacific) Language: Chinese Journal: Chinese Journal of Experimental Traditional Medical Formulae Year: 2022 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Language: Chinese Journal: Chinese Journal of Experimental Traditional Medical Formulae Year: 2022 Type: Article