Kazan Institute of Biochemistry and Biophysics, FRC Kazan Scientific Center of RAS, Kazan, Russia.
Methods Mol Biol. 2024;2791:35-43. doi: 10.1007/978-1-0716-3794-4_4.
The study of the localization of secondary metabolites in both plants and the cell cultures on the intravital sections is hampered by the difficulty of obtaining thin, correctly oriented sections. Techniques for fixing tissues in resins allow these difficulties to be overcome. Properly selected tissue fixation techniques allow using different dyes to identify the compound of interest. In addition, some components of tissue fixation can act as fixatives and as a dye for identifying secondary metabolites. For example, osmium tetroxide, which fixes lipids in tissues, stains phenolic compounds black. This paper describes methods for the detection of phenolic compounds in morphogenic callus culture of buckwheat using osmium tetroxide, Toluidine Blue O dye, and ferric chloride as dyes in epoxy resin-embedded cell culture with double fixation of the material and when material fixed in Karnovsky's fixative.
在活体切片上研究植物和细胞培养物中次生代谢物的定位受到难以获得薄而正确定向切片的阻碍。用于固定树脂中的组织的技术可以克服这些困难。适当选择的组织固定技术可以使用不同的染料来鉴定感兴趣的化合物。此外,组织固定的一些成分可以用作固定剂和鉴定次生代谢物的染料。例如,固定组织中脂质的四氧化锇将酚类化合物染成黑色。本文描述了使用锇四氧化物、甲苯胺蓝 O 染料和氯化铁作为染料,在环氧树脂包埋的细胞培养物中检测荞麦形态发生愈伤组织中酚类化合物的方法,其中材料经过双重固定,并且材料固定在 Karnovsky 固定剂中。