Horisberger M, Clerc M F
Nestlé Research Centre, Nestec Ltd., Lausanne, Switzerland.
Histochemistry. 1988;90(3):165-75. doi: 10.1007/BF00492504.
A new cationic colloidal gold complex has been developed for ultrastructural localization of cell surface anionic sites by transmission and scanning electron microscopy. The marker is prepared by labelling gold particles of suitable sizes (6 to 70 nm in diameter) with chitosan, a polymer of beta (1----4)-linked D-glucosamine. Using human red blood cells as a model, chitosan-gold complexes were shown to be specific for anionic sites and at pH 2 for sialic acid residues. The binding capacity of complexes of different sizes with carboxymethyl and phosphorylated celluloses was examined as a function of pH and ionic strength. The results indicated that these complexes can be used under acidic conditions as well as in physiological buffers. The complexes were further tested by transmission and scanning electron microscopy in detecting anionic sites on cells of various origins such as Escherichia coli, Lactobacillus maltaromicus, Lactobacillus reuteri, Saccharomyces cerevisiae, Saccharomyces rouxii, Schizosaccharomyces pombe, Fusarium oxysporum, Catharantus roseus.
已开发出一种新型阳离子胶体金复合物，用于通过透射电子显微镜和扫描电子显微镜对细胞表面阴离子位点进行超微结构定位。该标记物是通过用壳聚糖（一种β(1→4)连接的D-葡糖胺聚合物）标记合适尺寸（直径6至70纳米）的金颗粒制备而成。以人红细胞为模型，壳聚糖-金复合物显示对阴离子位点具有特异性，在pH 2时对唾液酸残基具有特异性。研究了不同尺寸的复合物与羧甲基纤维素和磷酸化纤维素的结合能力随pH和离子强度的变化。结果表明，这些复合物既可以在酸性条件下使用，也可以在生理缓冲液中使用。通过透射电子显微镜和扫描电子显微镜进一步测试了这些复合物在检测各种来源细胞（如大肠杆菌、麦芽糖乳杆菌、罗伊氏乳杆菌、酿酒酵母、鲁氏酵母、粟酒裂殖酵母、尖孢镰刀菌、长春花）表面阴离子位点方面的性能。
