Pearlmutter N L, Lembi C A
J Histochem Cytochem. 1978 Oct;26(10):782-91. doi: 10.1177/26.10.722047.
Chitin was visualized in cell walls after hydrolysis with potassium hydroxide and subsequent postfixation of the deacetylated polysaccharide (chitosan) in OsO4. Areas of chitin deposition appeared dark borwn by light microscopy and electron dense in the electron microscope. With this method, the presence of chitin was demonstrated in the cell walls of the green alga Pithophora oedogonia (Montagne) Wittrock and two fungi, Ceratocystis ulmi Buism. (C. Moreau) and Blastocladiella emersonii Cantino and Hyatt. Most of the chitin in P. oedogonia ws found in the crosswall disk and small amounts occurred in the outer longitudinal walls. The septal disk of C. ulmi also contained chitin, but significant amounts were present in the inner and outer regions of longitudinal walls as well. Chitin was present throughout the walls of B. emersonii. Small amounts of chitin were not easily demonstrated by this technique, but removal of chitosan by exposure to dilute acetic acid before osmium fixation disrupted cell wall integrity, suggesting that small amounts of the structural polysaccharide had been removed.
用氢氧化钾水解后，再用四氧化锇对脱乙酰化多糖（壳聚糖）进行后固定，可在细胞壁中观察到几丁质。通过光学显微镜观察，几丁质沉积区域呈深棕色，在电子显微镜下则表现为电子致密。用这种方法，在绿藻多列刚毛藻（Montagne）Wittrock以及两种真菌——榆枯萎病菌（Ceratocystis ulmi Buism. (C. Moreau)）和爱默生芽枝霉（Blastocladiella emersonii Cantino and Hyatt）的细胞壁中证实了几丁质的存在。多列刚毛藻中的大部分几丁质存在于横壁盘，少量存在于外部纵向壁中。榆枯萎病菌的隔膜盘也含有几丁质，纵向壁的内部和外部区域也有大量几丁质。几丁质存在于爱默生芽枝霉的整个细胞壁中。用这种技术不容易检测到少量几丁质，但在锇固定前用稀醋酸处理去除壳聚糖会破坏细胞壁的完整性，这表明少量的结构多糖已被去除。
