Foucrier J, Péchinot D, Feldmann G
J Immunol Methods. 1985 Dec 17;85(1):105-14. doi: 10.1016/0022-1759(85)90278-9.
A simple modification of the hemolytic plaque test is proposed which provides a morpho-functional approach to the study of secreting cells. The experimental procedure described here, an 'open system' with glass coverslips, involves the association of the reverse hemolytic plaque test performed in liquid medium with Feulgen's reaction used to reveal the nuclear characteristics of the secreting cells. A monolayer cell carpet, prepared with hepatocytes and sheep red blood cells coated with specific antibodies to transferrin, was attached to glass coverslips coated with 2 substrates, collagen and poly-L-lysine. Incubation was performed in a routine liquid culture medium. Under the conditions reported here to obtain an optimal attachment of the cells to the substrates, it was possible to stain hepatocytes after the formation of hemolytic plaques, whilst maintaining the monolayer cell structure intact. This technical modification should be of value in establishing correlations between secretory activity and cellular characteristics.
本文提出了一种溶血空斑试验的简单改良方法，该方法为分泌细胞的研究提供了一种形态功能学途径。这里描述的实验程序是一种使用玻璃盖玻片的“开放系统”，它涉及在液体培养基中进行的反向溶血空斑试验与用于揭示分泌细胞核特征的福尔根反应相结合。用肝细胞和包被有抗转铁蛋白特异性抗体的绵羊红细胞制备的单层细胞铺片，附着在包被有胶原蛋白和聚-L-赖氨酸这两种底物的玻璃盖玻片上。在常规液体培养基中进行孵育。在此报道的条件下，为使细胞最佳地附着于底物，溶血空斑形成后能够对肝细胞进行染色，同时保持单层细胞结构完整。这种技术改良在建立分泌活性与细胞特征之间的相关性方面应具有价值。
