Larsson L I, Hougaard D M
Department of Molecular Cell Biology, Statens Seruminstitut, Copenhagen S, Denmark.
Histochemistry. 1994 Jun;101(5):325-31. doi: 10.1007/BF00268993.
We have developed a model system based on the immobilization of microdroplets (0.1 microliters volumes) of antigens or of target (sense) oligonucleotides onto aminoalkylsilane-coated glass slides. Oligopeptide antigens need to be vapour-fixed in order to achieve efficient immobilization, while oligonucleotides do not require fixation. Protein antigens, exemplified by rabbit immunoglobulin, may be subjected to liquid fixation. The glass slide models are optically translucent and useful both for in situ hybridization and immunocytochemistry. The slides are compatible with detection systems of both light and fluorescence microscopy and permit measurement of staining intensities by microfluorimetry or computerized microdensitometry. The model systems can be used for comparisons of method sensitivities, for characterizing antibody and probe sensitivities and cross-reactivities, and as internal standards or quality controls for immunocytochemistry and in situ hybridization.
我们开发了一种模型系统,该系统基于将抗原或靶标(有义)寡核苷酸的微滴(体积为0.1微升)固定在氨基烷基硅烷包被的载玻片上。为了实现高效固定,寡肽抗原需要进行气相固定,而寡核苷酸则不需要固定。以兔免疫球蛋白为例的蛋白质抗原可进行液相固定。载玻片模型具有光学半透明性,可用于原位杂交和免疫细胞化学。这些载玻片与光学显微镜和荧光显微镜的检测系统兼容,并允许通过微量荧光测定法或计算机化显微密度测定法测量染色强度。该模型系统可用于比较方法的灵敏度,表征抗体和探针的灵敏度及交叉反应性,还可作为免疫细胞化学和原位杂交的内标或质量控制。