De Micco V, Aronne G
Laboratory of Botany and Reproductive Ecology, Department of Fruitculture, Botany and Plant Pathology, University of Naples Federico II, via Università, Portici, (NA), Italy.
Biotech Histochem. 2007 Aug;82(4-5):209-16. doi: 10.1080/10520290701713981.
Histological staining methods commonly used for detecting cellulose and lignin in cell walls were combined with epifluorescence microscopy to visualize differences in lignification between and within cellular elements. We tested our approach on sections of one-year-old branches of Fraxinus ornus L., Myrtus communis L., Olea europaea L., Pistacia lentiscus L. and Rhamnus alaternus L., containing both normal and tension wood. Sections were subjected to various staining techniques, viz. safranin O, safranin O/fast green FCF, and alcoholic solutions of safranin O/astra blue, according to the commonly accepted protocols. Stained and unstained sections were compared using both light and epifluorescence microscopy. Safranin O with or without counterstaining hid the strong fluorescence of vessel walls, cell corners and middle lamellae allowing the secondary wall fibers to fluoresce more clearly. Epifluorescence microscopy applied to stained sections showed more cell wall details than autofluorescence of unstained sections or white light microscopy of counterstained sections. This simple approach proved reliable and valuable for detecting differences in lignification in thick sections without the need for costly equipment.
常用于检测细胞壁中纤维素和木质素的组织学染色方法与落射荧光显微镜相结合，以观察细胞成分之间以及细胞内部木质化的差异。我们在白蜡树、香桃木、油橄榄、乳香黄连木和鼠李的一年生枝条切片上测试了我们的方法，这些枝条切片包含正常木材和应拉木。根据公认的方案，切片采用了各种染色技术，即番红O、番红O/固绿FCF以及番红O/阿斯特拉蓝酒精溶液。使用光学显微镜和落射荧光显微镜对染色和未染色切片进行比较。带有或不带有复染剂的番红O掩盖了导管壁、细胞角隅和胞间层的强荧光，使次生壁纤维发出更清晰的荧光。应用于染色切片的落射荧光显微镜比未染色切片的自发荧光或复染切片的白光显微镜显示出更多细胞壁细节。这种简单方法对于检测厚切片中木质化差异是可靠且有价值的，无需昂贵设备。
